Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Tadashi Ueda Last modified date:2021.10.27

Professor / Department of Medico-Pharmaceutical Scineces / Department of Pharmaceutical Health Care and Sciences / Faculty of Pharmaceutical Sciences

1. Takeru Ebihara , Akitsu Masuda , Daisuke Takahashi , Masato Hino , Hiroaki Mon, Kohei Kakino , Tsuguru Fujii , Ryosuke Fujita , Tadashi Ueda, Jae Man Lee , Takahiro Kusakabe, Production of scFv, Fab, and IgG of CR3022 Antibodies Against SARS-CoV-2 Using Silkworm-Baculovirus Expression System, Mol. Biotechnol., 63, 12, 1223-1234, 2021.12.
2. Kosuke Oyama, Takatoshi Ohkuri , Mao Inoue , Jose M M Caaveiro, Tadashi Ueda, High-level expression of human CH2 domain from the Fc region in Pichia pastoris and preparation of anti-CH2 antibodies, J Biochem, 10.1093/jb/mvab039, 2021.09.
3. Hitomi Nakamura , Moeka Yoshikawa , Naoko Oda-Ueda, Tadashi Ueda, Takatoshi Ohkuri, A comprehensive analysis of novel disulfide bond introduction site into the constant domain of human Fab, Sci Rep , 11, 1, 12937, 2021.06.
4. Kosuke Oyama, Takatoshi Ohkuri, Jinta Ochi, Jose M M Caaveiro, Tadashi Ueda, Abolition of aggregation of CH 2 domain of human IgG1 when combining glycosylation and protein stabilization, Biochem Biophys Res Commun., 10.1016/j.bbrc.2021.04.070, 558, 114-119, 2021.06.
5. Tatsuhiro Igawa , Shuhei Kishikawa, Yoshito Abe, Makoto Tsuda, Kazuhide Inoue, Tadashi Ueda, Analysis of binding residues in monoclonal antibody with high affinity for the head domain of the rat P2X4 receptor, J. Biochem. , 10.1093/jb/mvaa124, 169, 4, 491-496, 2021.04.
6. Hitomi Nakamura, Masato Kiyoshi , Makoto Anraku , Noritaka Hashii , Naoko Oda-Ueda , Tadashi Ueda , Takatoshi Ohkuri, Glycosylation decreases aggregation and immunogenicity of adalimumab Fab secreted from Pichia pastoris, J Biochem., 10.1093/jb/mvaa116, 169, 4, 435-443, 2021.04.
7. Yoshito Abe 􏰁, Yohei Ikeda, Saki Fujiyama, R. Manjunatha Kini, Tadashi Ueda,, A structural model of the PriB–DnaT complex in Escherichia coli replication restart, FEBS Letters, 595, 3, 341-350, 2021.02.
8. Yoshito Abe , Hinako Shibata , Kousuke Oyama, Tadashi Ueda , Effect of O-glycosylation on amyloid fibril formation of the variable domain in the Vλ6 light chain mutant Wil, Int J Biol Macromol ., 10.1016/j.ijbiomac.2020.10.194, 166, 342-351, 2021.01.
9. Hitomi Nakamura, Makoto Anraku, Naoko Oda-Ueda, Tadashi Ueda, Takatoshi Ohkuri, C-terminal cysteine PEGylation of adalimumab fab with an engineered interchain SS bond, Biological and Pharmaceutical Bulletin, 10.1248/bpb.b19-00612, 43, 3, 418-423, 2020.03, Conjugation with polyethylene glycol (PEG) is performed to increase serum half-life of the Fab for clinical applications. However, current designs for recombinant Fab only allow PEGylation at the interchain SS bond (disulfide bond) at the C-terminal end of the heavy chain and light chain of the Fab, which the decrease of thermostability occurred by partial reduction of the interchain SS bond. An adalimumab Fab mutant with a novel interchain SS bond (CH1:C177–CL:C160) and one cysteine at the C-terminal end (mutSS FabSH) was designed to maintain Fab thermostability and for site-specific PEGylation. MutSS FabSH was expressed in Pichia pastoris and purified mutSS FabSH was conjugated with 20-kDa PEG targeted at the free cysteine. Based on enzyme-linked immunosorbent assay (ELISA), PEGylation did not affect the binding capacity of the mutSS FabSH. To confirm the influence of PEGylation on the pharmacokinetic behavior of the Fab, PEGylated mutSS FabSH was administered to rats via tail vein injection. Analysis of the mean serum concentration of the PEGylated mutSS FabSH versus time through ELISA indicated an increase in half-life compared to that of non-PEGylated wild-type Fab. Consequently, we have successfully demonstrated that a Fab mutant with a novel interchain SS bond and one free cysteine at the C-terminal end can be PEGylated without changes in functionality. This design can potentially be used as a platform for modification of other recombinant Fabs..
10. Koji Nagata, Akitoshi Okada, Jun Ohtsuka, Takatoshi Ohkuri, Yusuke Akama, Yukari Sakiyama, Erika Miyazaki, Shoichiro Horita, Tsutomu Katayama, Tadashi Ueda, Masaru Tanokura, Crystal structure of the complex of the interaction domains of Escherichia coli DnaB helicase and DnaC helicase loader
Structural basis implying a distortion-accumulation mechanism for the DnaB ring opening caused by DnaC binding, Journal of biochemistry, 10.1093/jb/mvz087, 167, 1, 1-14, 2020.01, Loading the bacterial replicative helicase DnaB onto DNA requires a specific loader protein, DnaC/DnaI, which creates the loading-competent state by opening the DnaB hexameric ring. To understand the molecular mechanism by which DnaC/DnaI opens the DnaB ring, we solved 3.1-Å co-crystal structure of the interaction domains of Escherichia coli DnaB-DnaC. The structure reveals that one N-terminal domain (NTD) of DnaC interacts with both the linker helix of a DnaB molecule and the C-terminal domain (CTD) of the adjacent DnaB molecule by forming a three α-helix bundle, which fixes the relative orientation of the two adjacent DnaB CTDs. The importance of the intermolecular interface in the crystal structure was supported by the mutational data of DnaB and DnaC. Based on the crystal structure and other available information on DnaB-DnaC structures, we constructed a molecular model of the hexameric DnaB CTDs bound by six DnaC NTDs. This model suggested that the binding of a DnaC would cause a distortion in the hexameric ring of DnaB. This distortion of the DnaB ring might accumulate by the binding of up to six DnaC molecules, resulting in the DnaB ring to open..
11. Ohkuri Takatoshi, Yuge Natsuko, Sato Kenji, Ueda Tadashi, A method to induce hen egg lysozyme-specific humoral immune tolerance in mice by pre-exposition with the protein's oligomers., Biochemical and Biophysical Reports, doi: 10.1016/j.bbrep.2019.100679, 2019 Dec. 20:100679. eCollection, 2019.12.
12. Naoya Shindo, Hirokazu Fuchida, Mami Sato, Kosuke Watari , Tomohiro Shibata, Keiko Kuwata , Chizuru Miura , Kei Okamoto, Yuji Hatsuyama, Keisuke Tokunaga, Seiichi Sakamoto, Satoshi Morimoto, Yoshito Abe, Mitsunori Shiroishi, Jose M M Caaveiro, Tadashi Ueda , Tomonori Tamura , Naoya Matsunaga, Takaharu Nakao, Satoru Koyanagi, Shigehiro Ohdo, Yasuchika Yamaguch , Itaru Hamachi, Mayumi Ono, Akio Ojida, Selective and Reversible Modification of Kinase Cysteines With Chlorofluoroacetamides, Nature Chemical Biology, 10.1038/s41589-018-0204-3, 15, 3, 250-258, 2019.03.
13. Tatsuhiro Igawa, Shuhei Kishikawa, Yoshito Abe, Tomohiro Yamashita, Saki Nagai, Mitsunori Shiroishi, Chinatsu Shinozaki, Hiroyuki Tanaka, Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Kazuhide Inoue, Tadashi Ueda, Evidence for detection of rat P2X4 receptor expressed on cells by generating monoclonal antibodies recognizing the native structure, Purinergic Signalling, 10.1007/s11302-019-09646-5, 2019.01, P2X purinergic receptors are ATP-driven ionic channels expressed as trimers and showing various functions. A subtype, the P2X4 receptor present on microglial cells is highly involved in neuropathic pain. In this study, in order to prepare antibodies recognizing the native structure of rat P2X4 (rP2X4) receptor, we immunized mice with rP2X4's head domain (rHD, Gln111-Val167), which possesses an intact structure stabilized by S-S bond formation (Igawa and Abe et al. FEBS Lett. 2015), as an antigen. We generated five monoclonal antibodies with the ability to recognize the native structure of its head domain, stabilized by S-S bond formation. Site-directed mutagenesis revealed that Asn127 and Asp131 of the rHD, in which combination of these amino acid residues is only conserved in P2X4 receptor among P2X family, were closely involved in the interaction between rHD and these antibodies. We also demonstrated the antibodies obtained here could detect rP2X4 receptor expressed in 1321N1 human astrocytoma cells..
14. Saki Fujiyama, Yoshito Abe, Mitsunori Shiroishi, Yohei Ikeda, Tadashi Ueda, Insight into the interaction between PriB and DnaT on bacterial DNA replication restart
Significance of the residues on PriB dimer interface and highly acidic region on DnaT, Biochimica et Biophysica Acta - Proteins and Proteomics, 10.1016/j.bbapap.2019.01.008, 1867, 4, 367-375, 2019.01, When the replisome collapses at a DNA damage site, a sequence-independent replication restart system is required. In Escherichia coli, PriA, PriB, and DnaT assemble in an orderly fashion at the stalled replication fork and achieve the reloading of the replisome. PriB-DnaT interaction is considered a significant step in the replication restart. In this study, we examined the contribution of the residues Ser20, His26 and Ser55, which are located on the PriB dimer interface. These residues are proximal to Glu39 and Arg44, which are important for PriB-DnaT interaction. Mutational analyses revealed that His26 and Ser20 of PriB are important for the interaction with DnaT, and that the Ser55 residue of PriB might have a role in negatively regulating the DnaT binding. These residues are involved in not only the interaction between PriB and DnaT but also the dissociation of single-stranded DNA (ssDNA) from the PriB-ssDNA complex due to DnaT binding. Moreover, NMR study indicates that the region Asp66-Glu76 on the linker between DnaT domains is involved in the interaction with wild-type PriB. These findings provide significant information about the molecular mechanism underlying replication restart in bacteria..
15. Naoya Shindo, Hirokazu Fuchida, Mami Sato, Kosuke Watari, Tomohiro Shibata, Keiko Kuwata, Chizuru Miura, Kei Okamoto, Yuji Hatsuyama, Keisuke Tokunaga, Seiichi Sakamoto, Satoshi Morimoto, Yoshito Abe, Mitsunori Shiroishi, Jose M M Caaveiro, Tadashi Ueda, Tomonori Tamura, Naoya Matsunaga, Takaharu Nakao, Satoru Koyanagi, Shigehiro Ohdo, Yasuchika Yamaguchi, Itaru Hamachi, Mayumi Ono, Akio Ojida, Selective and reversible modification of kinase cysteines with chlorofluoroacetamides, Nature Chemical Biology, 10.1038/s41589-018-0204-3, 2019.01, Irreversible inhibition of disease-associated proteins with small molecules is a powerful approach for achieving increased and sustained pharmacological potency. Here, we introduce α-chlorofluoroacetamide (CFA) as a novel warhead of targeted covalent inhibitor (TCI). Despite weak intrinsic reactivity, CFA-appended quinazoline showed high reactivity toward Cys797 of epidermal growth factor receptor (EGFR). In cells, CFA-quinazoline showed higher target specificity for EGFR than the corresponding Michael acceptors in a wide concentration range (0.1-10 μM). The cysteine adduct of the CFA derivative was susceptible to hydrolysis and reversibly yielded intact thiol but was stable in solvent-sequestered ATP-binding pocket of EGFR. This environment-dependent hydrolysis can potentially reduce off-target protein modification by CFA-based drugs. Oral administration of CFA quinazoline NS-062 significantly suppressed tumor growth in a mouse xenograft model. Further, CFA-appended pyrazolopyrimidine irreversibly inhibited Bruton's tyrosine kinase with higher target specificity. These results demonstrate the utility of CFA as a new class warheads for TCI..
16. Yoshito Abe, Naoki Odawara, Nantanat Aeimhirunkailas, Hinako Shibata, Naoki Fujisaki, Hirofumi Tachibana, Tadashi Ueda, Inhibition of amyloid fibril formation in the variable domain of λ6 light chain mutant Wil caused by the interaction between its unfolded state and epigallocatechin-3-O-gallate, Biochimica et Biophysica Acta - General Subjects, 10.1016/j.bbagen.2018.08.006, 1862, 12, 2570-2578, 2018.12, Background: Light chains are abnormally overexpressed from disordered monoclonal B-cells and form amyloid fibrils, which are then deposited on the affected organ, leading to a form of systemic amyloidosis known as AL (Amyloid Light chain) amyloidosis. A green tea catechin, epigallocatechin-3-O-gallate (EGCG), which is thought to inhibit various amyloidoses, is a potent inhibitor of amyloid fibril formation in AL amyloidosis. Methods: An amyloidogenic variable domain in λ6 light chain mutant, Wil was incubated in the presence of EGCG. The incubation products were analyzed by SDS-PAGE and reverse-phase HPLC. The interaction between Wil and EGCG was observed by using NMR and tryptophan fluorescence. Results: EGCG inhibited the amyloid fibril formation of Wil at pH 7.5 and 42 °C. Under these conditions, most Wil populations were in the unfolded state and several chemical reactions, i.e., oxidation and/or covalent bond oligomerization could be induced by auto-oxidated EGCG. Moreover, we found that EGCG bound to the unfolded state of Wil with higher affinity (Kd = 7 μM). Conclusions: Inhibition of amyloid fibril formation of Wil was caused by 1) EGCG binding to unfolded state rather than folded state and 2) chemical modifications of Wil by auto oxidation of EGCG. General significance: In the competitive formation of amyloid fibrils and off-pathway oligomers, EGCG produces the latter immediately after it preferentially binds to the unfolded state. It may be general mechanism of EGCG inhibition for amyloidosis..
17. Hitomi Nakamura, Naoko Oda-Ueda, Tadashi Ueda, Takatoshi Ohkuri, Introduction of a glycosylation site in the constant region decreases the aggregation of adalimumab Fab, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2018.06.071, 503, 2, 752-756, 2018.09, The production of therapeutic monoclonal antibodies is costly; therefore, antigen-binding fragments (Fabs) can be used instead. However, their tendency toward aggregation can reduce the half-life in the plasma and the therapeutic effectiveness. To examine the effect of glycosylation on the properties of the Fab of a therapeutic antibody, an N-glycosylation site was introduced at position 178 of the H-chain constant region of adalimumab Fab through site-directed mutagenesis of L178 N (H:L178 N Fab), and then H:L178 N Fab was expressed in Pichia pastoris. SDS-PAGE analysis with treatment of N-glycosidase F or periodic acid–Schiff reagent showed that H:L178 N Fab contained a relatively low glycan level. Moreover, the H:L178 N mutation did not decrease the binding activity and thermal stability of Fab, and H:L178 N Fab was more resistant to protease digestion than wild-type Fab. The aggregation of Fab induced by pH-shift stress was measured by monitoring the optical density at 350 nm. Although the wild-type Fab showed a large increase in optical density with an increase of protein concentration, no such increase of turbidity during aggregation was found in H:L178 N Fab. These results demonstrated that glycosylation at position 178 of the H-chain constant region of adalimumab Fab can prevent protein aggregation, and therefore serve as a potentially effective platform for drug development..
18. Kazuhiro Miyanabe, Takefumi Yamashita, Yoshito Abe, Hiroki Akiba, Yuichiro Takamatsu, Makoto Nakakido, Takao Hamakubo, Tadashi Ueda, Jose Manuel Martinez Caaveiro, Kouhei Tsumoto, Tyrosine Sulfation Restricts the Conformational Ensemble of a Flexible Peptide, Strengthening the Binding Affinity for an Antibody, Biochemistry, 10.1021/acs.biochem.8b00592, 57, 28, 4177-4185, 2018.07, Protein tyrosine sulfation (PTS) is a post-translational modification regulating numerous biological events. PTS generally occurs at flexible regions of proteins, enhancing intermolecular interactions between proteins. Because of the high flexibility associated with the regions where PTS is generally encountered, an atomic-level understanding has been difficult to achieve by X-ray crystallography or nuclear magnetic resonance techniques. In this study, we focused on the conformational behavior of a flexible sulfated peptide and its interaction with an antibody. Molecular dynamics simulations and thermodynamic analysis indicated that PTS reduced the main-chain fluctuations upon the appearance of sulfate-mediated intramolecular H-bonds. Collectively, our data suggested that one of the mechanisms by which PTS may enhance protein-protein interactions consists of the limitation of conformational dynamics in the unbound state, thus reducing the loss of entropy upon binding and boosting the affinity for its partner..
19. Hitomi Nakamura, Naoko Oda-Ueda, Tadashi Ueda, Takatoshi Ohkuri, A novel engineered interchain disulfide bond in the constant region enhances the thermostability of adalimumab Fab, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.10.140, 495, 1, 7-11, 2018.01, We constructed a system for expressing the Fab of the therapeutic human monoclonal antibody adalimumab at a yield of 20 mg/L in the methylotrophic yeast Pichia pastoris. To examine the contribution of interchain disulfide bonds to conformational stability, we prepared adalimumab Fab from which the interchain disulfide bond at the C-terminal region at both the CH1 and CL domains was deleted by substitution of Cys with Ala (FabΔSS). DSC measurements showed that the Tm values of FabΔSS were approximately 5 °C lower than those of wild-type Fab, suggesting that the interchain disulfide bond contributes to conformational thermostability. Using computer simulations, we designed a novel interchain disulfide bond outside the C-terminal region to increase the stability of FabΔSS. The resulting Fab (mutSS FabΔSS) had the mutations H:V177C and L:Q160C in FabΔSS, confirming the formation of the disulfide bond between CH1 and CL. The thermostability of mutSS FabΔSS was approximately 5 °C higher than that of FabΔSS. Therefore, the introduction of the designed interchain disulfide bond enhanced the thermostability of FabΔSS and mitigated the destabilization caused by partial reduction of the interchain disulfide bond at the C-terminal region, which occurs in site-specific modification such as PEGylation..
20. Mitsunori Shiroishi, Yuji Ito, Kenta Shimokawa, Man Lee, Takahiro Kusakabe, Tadashi Ueda, Structure–function analyses of a stereotypic rheumatoid factor unravel the structural basis for germline-encoded antibody autoreactivity, Journal of Biological Chemistry, 10.1074/jbc.M117.814475, 293, 18, 7008-7016, 2018.01, Rheumatoid factors (RFs) are autoantibodies against the fragment-crystallizable (Fc) region of IgG. In individuals with hematological diseases such as cryoglobulinemia and certain B cell lymphoma forms, the RFs derived from specific heavy- and light-chain germline pairs, so-called “stereotypic RFs,” are frequently produced in copious amounts and form immune complexes with IgG in serum. Of note, many structural details of the antigen recognition mechanisms in RFs are unclear. Here we report the crystal structure of the RF YES8c derived from the IGHV1-69/IGKV3-20 germline pair, the most common of the stereotypic RFs, in complex with human IgG1-Fc at 2.8 Å resolution. We observed that YES8c binds to the CH2–CH3 elbow in the canonical antigen-binding manner involving a large antigen–antibody interface. On the basis of this observation, combined with mutational analyses, we propose a recognition mechanism common to IGHV1-69/IGKV3-20 RFs: (1) the interaction of the Leu
region of Fc enables the highly variable complementarity-determining region (CDR)-H3 to participate in the binding, (2) the hydrophobic tip in the CDR-H2 typical of IGHV1-69 antibodies recognizes the hydrophobic patch on Fc, and (3) the interaction of the highly conserved RF light chain with Fc is important for RF activity. These features may determine the putative epitope common to the IGHV1-69/IGKV3-20 RFs. We also showed that some mutations in the binding site of RF increase the affinity to Fc, which may aggravate hematological diseases. Our findings unravel the structural basis for germline-encoded antibody autoreactivity..
21. Shohei Mine, Masahiro Watanabe, Saori Kamachi, Yoshito Abe, Tadashi Ueda, The structure of an archaeal β-glucosaminidase provides insight into glycoside hydrolase evolution., J. Biol. Chem., 10.1074/jbc.M116.766535., 292, 12, 4996-5006, 2017.01.
22. Hitomi Nakamura, Takatoshi Ohkuri, Takanori So, Tadashi Ueda, Relationship between the magnitude of IgE production in mice and conformational stability of the house dust mite allergen, Der p 2., Biochimica et Biophysica Acta (BBA) - General Subjects, 10.1016/j.bbagen, in press, 2016.06.
23. Yoshito Abe, Mitsuru Kubota, Shinta Takazaki, Yuji Ito, Hiromi Yamamoto, Dongchon Kang, Tadashi Ueda, Taiji Imoto, Effect on catalysis by replacement of catalytic residue from hen egg white lysozyme to Venerupis philippinarum lysozyme, Protein Science, 10.1002/pro.2966, 25, 9, 1637-1647, 2016.06.
24. Yoshito Abe, Naoki Fujisaki, Takanori Miyoshi, Noriko Watanabe, Tsutomu Katayama, Tadashi Ueda, Functional analysis of CedA based on its structure: residues important in binding of DNA and RNA polymerase and in the cell division regulation, The Journal of Biochemistry, 10.1093/jb/mvv096, 159, 2, 217-223, 2016.02.
25. Takahiro Aramaki, Yoshito Abe, Kaori Furutani, Tsutomu Katayama, Tadashi Ueda, Basic and aromatic residues in the C-terminal domain of PriC are involved in ssDNA and SSB binding., J. Biochem., 10.1093/jb/mvv014., 157, 6, 529-537, 2015.06.
26. Yoshito Abe, Takatoshi Ohkuri, Sachiko Yoshitomi, Tadashi Ueda, Role of the osmolyte taurine on the folding of a model protein, hen egg white lysozyme, under a crowding condition, AMINO ACIDS, 10.1007/s00726-015-1918-0, 47, 5, 909-915, 2015.05.
27. Tatsuhiro Igawa, Yoshito Abe, TSUDA MAKOTO, Kazuhide Inoue, Tadashi Ueda, Solution structure of the rat P2X4 receptor head domain involved in inhibitory metal binding, FEBS LETTERS, 10.1016/j.febslet.2015.01.034, 589, 6, 680-686, 2015.03.
28. Saki Fujiyama, Yoshito Abe, Junya Tani, Masashi Urabe, Kenji Sato, Takahiko Aramaki, Tsutomu Katayama, Tadashi Ueda, Structure and mechanism of the primosome protein DnaT-functional structures for homotrimerization, dissociation of ssDNA from the PriB.ssDNA complex, and formation of the DnaT.ssDNA complex, FEBS JOURNAL, 10.1111/febs.13080, 281, 23, 5356-5370, 2014.12.
29. Shohei Mine, Yuji Kado, Masahiro Watanabe, Yohta Fukuda, Yoshito Abe, Tadashi Ueda, Yutaka Kawarabayashi, Tsuyoshi Inoue, Kazuhiko Ishikawa, The structure of hyperthermophilic β-N-acetylglucosaminidase reveals a novel dimer architecture associated with the active site, FEBS journal, 10.1111/febs.13049, 281, 22, 5092-5103, 2014.08.
30. Shouhei Mine, Mayumi Niiyama, Wakana Hashimoto, Takahisa Watanabe, Daisuke Koma, Takashi Ohmoto, Yohta Fukuda, Tsuyoshi Inoue, Yoshito Abe, Tadashi Ueda, Junji Morita, Koichi Uegaki, Tsutomu Nakamura, Expression from engineered Escherichia coli chromosome and crystallographic study of archaeal N,N-diacetylchitobiose deacetylase, 10.1111/febs.12805. , 281, 11, 2584-2596, 2014.06.
31. Saki Fujiyama, Yoshito Abe, Taiichi Takenawa, Takahiko Aramaki, Seijiro Shioi, Tsutomu Katayama, Tadashi Ueda, Involvement of histidine in complex formation of PriB and single-stranded DNA, BBA-proteins and proteomics, 1844, 2, 299-307, 2014.02.
32. Masayuki Su'etsugu, Yuji Harada, Kenji Keyamura, Chika Matsunaga, Kazutoshi Kasho, Yoshito Abe, Tadashi Ueda, Tsutomu Katayama, The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA, Environ Microbiol., 15, 12, 3183-3195, 2013.12.
33. Takatoshi Ohkuri, Eri Murase, Sun Shu-Lan, Jun Sugitani, Tadashi Ueda, Characterization of deamidation at Asn138 in L-chain of recombinant humanized Fab expressed from Pichia pastoris, J.Biochem, 154, 4, 333-340, 2013.10.
34. Takahiko Aramaki, Yoshito Abe, Takatoshi Ohkuri, Tomonori Mishima, Shoji Yamashita, Tsutomu Katayama, Tadashi Ueda, Domain separation and characterization of PriC, a replication restart primosome factor in Escherichia coli. , Genes to Cells, 18, 9, 723-732, 2013.09.
35. Takahiko Aramaki, Yoshito Abe, Tsutomu Katayama, Tadashi Ueda, Solution structure of the N-terminal domain of a replication restart primosome factor, PriC, in Escherichia coli., Protein Science, 22, 9, 1279-1286, 2013.09.
36. Masato Abe, Yoshito Abe, Takaoshi Ohkuri, Tomonori Mishima, Akira Monji, Shigenobu Kanba, Tadashi Ueda, Mechanism for retardation of amyloid fibril formation by sugars in Vλ6 protein., Protein Science, 22,4,467-474, 4, 467-474, 2013.04.
37. Tatsuhiro Igawa, Sadayuki Higashi, Yoshito Abe, Takatoshi Ohkuri, Hiroyuki Tanaka, satoshi morimoto, 山下 智大, TSUDA MAKOTO, Kazuhide Inoue, Tadashi Ueda, Preparation and characterization of a monoclonal antibody against the refolded and functional extracellular domain of rat P2X4 receptor, J. Biochem., 153, 3, 275-282, 2013.03.
38. Shinya Takazaki, Yoshito Abe, Tomohiro Yamaguchi, Mikako Yagi, Tadashi Ueda, Dongchon Kang, Naotaka Hakasaki, C-terminal tail is involved in conformational change but not in substrate binding, BBA – Biomembranes, 1818, 3, 658-665, 2013.03.
39. Takazaki S, Abe Y, Yamaguchi T, Yagi M, Ueda T, Kang D, Hamasaki N., Arg 901 in the AE1 C-terminal tail is involved in conformational change but not in substrate binding, BBA – Biomembranes, 1818, 3, 658, 2012.03.
40. Kameoka D, Ueda T, Imoto T., Effect of the Conformational Stability of the CH2 Domain on the Aggregation and Peptide Cleavage of a Humanized IgG., Appl Biochem Biotechnol., 164, 5, 642, 2011.07.
41. Ohkuri T, Nagatomo S, Oda K, So T, Imoto T, Ueda T, A Protein's conformational stability is an immunolgically dominant factor: Evidence that free-energy barriers for protein unfolding limit the immunolgenicity of foreign proteins, The Journal of Immunlogy, 185, 4199-4205, 2010.10, 蛋白質は立体構造を保持した状態でその機能を発揮する。蛋白質は往々にして、その安定性を高めることで有用性が上がることが知られている。一方、生体内で蛋白質抗原は、立体構造が壊れた状態で細胞内プロテアーゼにより分解され抗原性を発揮する。従って、免疫応答にも蛋白質の安定性が関与するということを示唆する知見は得られていた。本報告では、蛋白質抗原が免疫応答を引き起こす主要な要因の一つは、蛋白質抗原の構造安定性であることを世界で初めて実証した。この研究成果は,最近世界的に使用が広がっている抗体医薬品の機能を高めるためにアミノ酸配列を改変したとしても、不測の免疫応答を回避する方策を示唆したものである。この成果によって、今後、高機能化蛋白質医薬品の研究が一層進展することが期待される.
42. Horikawa H, Kato TA, Mizoguchi Y, Monji A, Seki Y, Ohkuri T, Gotoh L, Yonaha M, Ueda T, Hashioka S, Kanba S, Inhibitory effects of SSRIs on INF-gamma induced microglial activation through the regulation of intracellular calcium, Prog. Neurophsychopharmacol. Biol. Psychiatry, 34, 1306-1316, 2010.06.
43. Goto T, Abe Y, Imoto T, Ueda T, Effect of protein concentration and pH on the chitinase activity of Tapes japonica lysozyme , Protein Pepti. lett., 2010.02.
44. Nagata-Uchiyama M, Abe Y, Monji A, Kanba S, Ueda T, Evidence for the binding of phosphate ion to the C-terminus region in Aβ1-40 using heteronuclear NMR analyses , Protein Pept. lett., 2010.01.
45. Tomonori Mishima, Takatoshi Ohkuri, Akira Monji, Takaaki Kanemaru, Yoshito Abe, Tadashi Ueda, Effect of His mutantions on the fibrillation of amyloidogenid Vλ6 protein Wil under acidic and physicological conditions, Biochemical and Biophysical Research Communication, 391, 1, 615, 2010.01.
46. Mishima T, Ohkuri T, Monji A, Kanemaru T, Abe Y, Ueda T, Residual Structures in the Acid-Unfolded States of Vlambda6 Proteins Affect Amyloid Fibrillation, J Mol Biol, 392(4):1033-43, 2009.10.
47. Keyamura K, Abe Y, Higashi M, Ueda T, Katayama T., DiaA dynamics are coupled with changes in initial origin complexes leading to helicase loading., J Biol Chem, 284(37):25038-50, 2009.09.
48. Ueda Y, Ohwada S, Abe Y, Shibata T, Iijima M, Yoshimitsu Y, Koshiba T, Nakata M, Ueda T, Kawabata SI., Factor G Utilizes a Carbohydrate-Binding Cleft That Is Conserved between Horseshoe Crab and Bacteria for the Recognition of {beta}-1,3-d-Glucan, J Immnol, 183(6):3810-8, 2009.09.
49. Takashi Goto, Takatoshi Ohkuri, Seijiro Shioi, Yoshito Abe, Taiji Imoto and Tadashi Ueda, Crystal Structure of K33 Mutant Hen Lysozymes with Enhanced Activities, The Journal of Biochemistry, Vol.144、No.5、619-623, 2008.11.
50. Ohkuri T, Takeda C, Yoshida Y, Izuhara K, Imoto T, Ueda T, Expression of human IL-13 receptor alpha2 extracellular domain in Pichia pastoris, Protein Expr Purif., 56、1、48, 2007.11.
51. Kameoka D, Matsuzaki E, Ueda T, Imoto T, Effect of buffer species on the unfolding and the aggregation of humanized IgG, The Journal of Biochemistry, 142、3、383, 2007.09.
52. Goto T, Abe Y, Kakuta Y, Takeshita K, Imoto T, Ueda T, Crystal structure of Tapes japonica lysozyme with substrate analogue; Structural basis of the catalytic mechanism and manifestation of its chitinase activity accompany with quaternary structural change, Jounal of Biological Chemistry, 282、37、27459, 2007.09.
53. Makiko Nagata-Uchiyama, Masashi Yaguchi, Yugo Hirano and Tadashi Ueda, Expression and Purification of Uniformly 15N-Labeled Amyloid beta Peptide 1-40 in Escherichia coli, Protein & Peptide Letters, 14, 8, 788-792, 2007.09.
54. Mishima T, Ohkuri T, Imoto T, Ueda T, Analyses of native disulfide bond formation in the early stage of the folding process in mutant lysozymes where the long-range interactions in the denatured state were modulated, Biosci. Biotechnol. Biochem., 71、8、2072, 2007.08.
55. Abe Y, Jo T, Matsuda Y, Matsunaga C, Katayama T, Ueda T, Structure and function of DnaA N-terminal domains: Specific sites and mechanisms in inter-DnaA interaction and in DnaB helicase loading on Ori C, Jounal of Biological Chemistry, 282、24、17816-17827, 2007.06.
56. Yoshida Y, Ohkuri T, Takeda C, Kuroki R, Izuhara K, Imoto T, Ueda T, Analysis of internal motions of interleukin 13 variant associated with severe bronchial asthma using 15N relaxation measurements, Biochem. Biophys. Res. Commun., 358、1、292-297, 2007.06.
57. Mishima T, Ohkuri T, Monji A, Imoto T, Ueda T, A paticular hydrophobic cluster in the residual structure of reduced lysozyme drastically affects the amyloid fibrils formation, Biochem. Biophys. Res. Commun., 356, 3, 769-772, 2007.05.
58. Fujii T, Ohkuri T, Onodera R, Ueda T, Stable supply of large amounts of human Fab from the inclusion bodies in E. coli., The Journal of Biochemistry, 141、5、699-707, 2007.05.
59. Abe Y, Watanabe N, Yoshida Y, Ebata F, Katayama T, Ueda T, Assignment of 1H, 13C and 15N resonances of N-terminal Domain of DnaA protein, Biomolecular NMR Assignments, 印刷中(当該雑誌Webで公開中), 2007.05.
60. Mishima T, Ohkuri T, Monji A, Imoto T, Ueda T, Amyloid formation in denatured single-mutant lysozyme where residual structures are modulated, Protein Science, 15, 10, 2448-2452, 2006.10.
61. Yoshida Y, Tanaka M, Ohkuri t, Tanaka Y, Imoto T, Ueda T, Analysis of internal motion of RNase T1 complexed with a productive substrate involving 15N NMR relaxation measurements, The Journal of Biochemistry, 140, 1, 43-48, 2006.07.
62. Ohkuri T, Imoto T, Ueda T, Effect of W62G mutation of hen lysozyme on the folding in vivo, Biochem. Biophys Res. Commun., 10.1016/j.bbrc.2005.10.009, 338, 2, 820-824, 338, 2, 820-824, 2005.12.
63. Hashioka S, Monji A, Ueda T, Kanba S, Nakanishi H, Amyloid-beta fibril formation is not necessarily required for microglial activation by the peptides, Neurochem Int., 10.1016/j.neuint.2005.05.001, 47, 5, 369-376, 47, 5, 369-376, 2005.10.
64. Ohkuri T, Imoto T, Ueda T, Characterization of refolded hen lysozyme variant lacking outside two disulfide bonds, Biosci. Biotechnol. Biochem., 10.1271/bbb.69.1206, 69, 6, 1206-1208, 69, 6, 1206-1208, 2005.06.
65. Yoshida Y, Ohkuri T, Kino T, Ueda T and Imoto T, Elucidation of the relationship between enzyme activity and internal motion using a lysozyme stabilized by cavity-filling mutations, Cellular and Molecular Life Sciences, 10.1007/s00018-005-5053-z, 62, 9, 1047-1055, 62, 5, 1047-1055, 2005.05.
66. Ohkuri T, Shioi S, Imoto T, Ueda T, Effect of the structure of the denatured state of lysozyme on the aggregation reaction at the early stages of folding from the reduced form, J. Mol. Biol., 10.1016/j.jmb.2005.01.022, 347, 1, 159-168, 347、1、159-168, 2005.03.
67. Ueno K, Ueda T, Sakai K, Abe Y, Hamasaki N, Okamoto M, Imoto T, Evidence for a novel racemization process of an asparaginyl residue in mouse lysozyme under physiological conditions, Cellular and Molecular Life Sciences, 10.1007/s00018-004-4412-5, 62, 2, 199-205, 62、2、199-205, 2005.01.
68. Shioi S, Ose T, Maenaka K, Shiroishi M, Abe Y, Kohda D, Katayama T, Ueda T, Crystal structure of a biologically functional form of PriB from Escherichia coli reveals a potential single-stranded DNA-binding site, Biochem. Biophys. Res. Commun., 10.1016/j.bbrc.2004.11.104, 326, 4, 766-776, 326、4、766-776, 2005.01.
69. Wirmer J, Schlorb C, Klein-Seetharaman J, Hirano R, Ueda T, Imoto T, Schwalbe H, Modulation of compactness and long-range interactions of unfolded lysozyme by single point mutations, Angewandte Chemie-International Edition, 10.1002/anie.200460907, 43, 43, 5780-5785, 43、43、5780-5785, 2004.11.
70. Shioi S, Imoto T, Ueda T, Analysis of the early stage of the folding process of reduced lysozyme using all lysozyme variants containing a pair of cysteines, Biochemistry, 10.1021/bi036048h, 43, 18, 5488-5493, 43、18、5488-5493, 2004.05.
71. Takeshita K, Hashimoto Y, Ueda T, Imoto T., A small chimerically bifunctional monomeric protein: Tapes japonica lysozyme., Cell Mol Life Sci., 10.1007/s00018-003-3082-z, 60, 9, 1944-1951, 60, 9, 1944-1951, 2003.09.
72. Yoshida Y, Obita T, Kokusho Y, Ohmura T, Katayama T, Ueda T, Imoto T., Identification of the region in Escherichia coli DnaA protein required for specific recognition of the DnaA box., Cell Mol Life Sci., 10.1007/s00018-003-3176-7, 60, 9, 1998-2008, 60, 9, 1998-2008, 2003.09.
73. Kameoka D, Ueda T, Imoto T., Method for the Detection of Asparagine Deamidation and Aspartate Isomerization of Proteins by MALDI/TOF-Mass Spectrometry Using Endoproteinase Asp-N., J Biochem., 134, 1, 129-135, 2003.07.
74. Numata T, Suzuki A, Kakuta Y, Kimura K, Yao M, Tanaka I, Yoshida Y, Ueda T, Kimura M., Crystal structures of the ribonuclease MC1 mutants N71T and N71S in complex with 5'-GMP: structural basis for alterations in substrate specificity., Biochemistry, 10.1021/bi034103g, 42, 18, 5270-5278, 42, 18, 5270-5278, 2003.04.
75. Obita T. Ueda T. Imoto T., Solution structure and activity of mouse lysozyme M., Cell. Mol. Life Sci., 10.1007/s000180300012, 60, 1, 176-184, 60, 1, 176-184, 2003.01.
76. Obita T. Iwura T. Su'etsugu M. Yoshida Y. Tanaka Y. Kayatama T. Ueda T. Imoto T., Determination of the secondary structure in solution of the Escherichia coli DnaA DNA-binding domain., Biochem. Biophys. Res. Commun., 10.1016/S0006-291X(02)02590-1, 299, 1, 42-48, 299, 1, 42-48, 2002.11.
77. Klein-Seetharaman J, Oikawa M, Grimshaw S B, Wirmer J, Durchardt E, Ueda T.他4名, Long-range interactions within a nonnative protein., Science, 10.1126/science.1067680, 295, 5560, 1719-1722, 295, 5560, 1719-1722, 2002.03.
78. Nishida S. Fujimitsu K. Sekimizu K. Ohmura T. Ueda T. Katayama T., A nucleotide switch in the E. coli DnaA protein initiates chromosomal replication: Evidence from a mutant DnaA protein defective in regulatory ATP hydrolysis in vitro and in vivo., J. Biol. Chem., 10.1074/jbc.M108303200, 277, 17, 14986-14995, 277, 17, 14986-14995, 2002.02.
79. Ohkuri T. Ueda T. Yoshida Y. Abe Y. Hamasaki N. Imoto T., A metal binding in the polypeptide chain improves the folding efficiency of a denatured and reduced protein., Biopolymers, 10.1002/bip.10153, 64, 2, 106-114, 642, 2, 106-114, 2002.02.
80. Masumoto K. Ueda T. Nagata M. Yamada Y. Yoshida Y. Hashimoto Y. Imoto T., Effects of stereochemistry of sugars on protein stabilities., Protein Peptide Lett., 10.2174/0929866023408553, 9, 5, 435-439, 9, , 435-439, 2002.01.
81. Monji A. Utsumi H. Ueda T. Imoto T. Yoshida I. Hashioka S. Tashiro K. Tashiro N., Amyloid-beta-protein (Ab) (25-35) - associated free radical generation is strongly influenced by the aggregational state of the peptides., Life Sciences, 10.1016/S0024-3205(01)01451-5, 70, 7, 833-841, 70, , 833-841, 2002.01.
82. Ueda T. Nagata M. Monji A. Yoshida I. Tashiro N. Imoto T., Effect of sucrose on formation of the beta-amyloid fibrils and D-aspartic acids in Ab 1-42., Biol. Pharm. Bull., 10.1248/bpb.25.375, 25, 3, 375-378, 25, , 75-378, 2002.01.
83. Ohmura T. Motoshima H. Ueda T. Imoto T., Fluctuations in free or substrate-complexed lysozyme and a mutant of it detected on x-ray crystallography and comparison with those detected on NMR., J. Biochem., 131, 5, 701-704, 131, , 701-704, 2002.01.
84. Tsujihata Y. Mizukami Y. Ueda T. So T. Imoto T., B-cell repertoire specific for an unfolded self-determinant of mouse lysozyme escape tolerance and dominantly participate in the autoantibody response., Immunology, 10.1046/j.1365-2567.2002.01528.x, 107, 4, 394-402, 107, , 394-402, 2002.01.
85. Ohmura T. Ueda T. Ootsuka K. Saito M. Imoto T., Stabilization of hen egg white lysozyme by a cavity-filling mutation., Protein Sci., 10.1110/ps.37401, 10, 2, 313-320, 10, 9, 313-320, 2001.09.
86. Ohmura T. Ueda T. Hashimoto Y. Imoto T., Tolerance of point substitution of methionine for isoleucine in hen egg white lysozyme., Protein Eng., 10.1093/protein/14.6.421, 14, 6, 421-425, 14, 6, 421-425, 2001.06.
87. Monji A. Utsumi H. Ueda T. Imoto T. Yoshida I. Hashioka S. Tashiro K. Tashiro N., The relationship between the aggregational state of the amyloid-beta peptides and free radical generation by the peptides. J. Neurochem. 77, 1425-1432, J. Neurochem., 10.1046/j.1471-4159.2001.00392.x, 77, 6, 1425-1432, 2001.01.
88. Hashimoto Y. Munemura O. Masumoto K. Ueda T. Imoto T., Thermostability of doubly glycosylated recombinant lysozyme., Biol. Pharm. Bull., 10.1248/bpb.24.1102, 24, 10, 1102-1107, 24, , 1102-1107, 2001.01.
89. Tsujihata Y. So T. Hashimoto Y. Ueda T. Imoto T., A single amino acid substitution in a self protein is sufficient to trigger autoantibody response., Mol. Immunol., 10.1016/S0161-5890(01)00068-2, 38, 5, 375-381, 38, , 375-381, 2001.01.
90. Ohkuri T. Ueda T. Tsurumaru M. Imoto T., Evidence for an initiation site for hen lysozyme folding from reduced form using its dissected peptide fragments., Protein Eng., 10.1093/protein/14.11.829, 14, 11, 829-833, 14, , 829-833, 2001.01.
91. Ueda T. Nagata M. Imoto T., Aggregation and chemical reaction in hen lysozyme caused by heating at pH 6 are depressed by osmolytes, sucrose and trehalose., J. Biochem., 130, 4, 491-496, 130, ,491-496, 2001.01.
92. So T. Ito H. Hirata M. Ueda T. Imoto T., Contribution of conformational stability of hen lysozyme to induction of type 2 T-helper immune responses., Immunology, 10.1046/j.1365-2567.2001.01314.x, 104, 3, 259-268, 104, , 289-268, 2001.01.
93. Mine S. Ueda T. Hashimoto Y. Imoto T., Analysis of the internal motion of free and ligand-bound human lysozyme by use of N-15 NMR relaxation measurement: A comparison with those of hen lysozyme., Protein Sci., 9, 9, 1669-1684, 9, 9,1669-1684, 2000.09.
94. Obita T. Ueda T. Tanaka Y. Hashimoto Y. Imoto T., Assignment of 1H and 15N resonances of mouse lysozyme M., J. Biomol. NMR, 18, 4, 361-362, 2000.04.
95. Masumoto K. Ueda T. Motoshima H. Imoto T., The relationship between local structure and stability in hen egg white lysozyme mutant with alanine substituted for glycine., Protein Eng., 13, , 691-695, 2000.01.
96. Hashimoto Y. Ikenaga T. Tanigawa K. Ueda T. Ezaki I. Imoto T., Expression and characterization of human rheumatoid factor single-chain Fv., Biol. Pharm. Bull., 23, 8, 941-945, 23, , 941-945, 2000.01.
97. Tsujihata Y. So T. Chijiiwa Y. Hashimoto Y. Hirata M. Ueda T. Imoto T., Mutant mouse lysozyme carrying a minimal T cell epitope of hen egg lysozyme evokes high autoantibody, J. Immunol., 165, 7, 3606-3611, 165, 3, 606-3611, 2000.01.
98. Ueda T. Masumoto K. Ishibashi R. So T. Imoto T., Remarkable thermal stability of doubly intramolecularly cross-linked hen lysozyme., Protein Eng., 10.1093/protein/13.3.193, 13, 3, 193-196, 13, , 193-196, 2000.01.
99. Mine S, Tate S, Ueda T, Kainosho M, Imoto T, Analysis of the relationship between enzyme activity and its internal motion using nuclear magnetic resonance:15N relaxation studies of wild-type and mutant lysozyme, Journal of Molecular Biology, 10.1006/jmbi.1999.2572, 286, 5, 1547-1565, 285、5、1547-1565, 1999.02.
100. Fujita H. Saeki M. Yasunaga K. Ueda T. Imoto T. Himeno M., In vitro binding study of adaptor protein complex (AP-1) to lysosomal targeting motif (LI-motif)., Biochem. Biophys. Res. Commun., 10.1006/bbrc.1998.0140, 255, 1, 54-58, 255, , 54-58, 1999.01.
101. Hashimoto Y. Tanigawa K. Nakashima M. Sonoda K. Ueda T. Watanabe T. Imoto T., Construction of the single-chain Fv from 196-14 antibody toward ovarian cancer-associated antigen CA125., Biol. Pharm. Bull., 22, 10, 1068-1072, 22, , 1068-1072, 1999.01.
102. So T. Ito HO. Tsujihata Y. Hirata M. Ueda T. Imoto T., The molecular weight ratio of monomethoxypolyethylene glycol (mPEG) to protein determines the immunotolerogenicity of mPEG proteins., Protein Eng., 10.1093/protein/12.8.701, 12, 8, 701-705, 12, ,701-705, 1999.01.
103. So T. Ito HO. Hirata M. Ueda T. Imoto T., Extended blood half-life of monomethoxypolyethylene glycol-conjugated hen lysozyme is a key parameter controlling immunological tolerogenicity., Cell. Mol. Life Sci., 10.1007/s000180050365, 55, 8-9, 1187-1194, 55, , 1187-1194, 1999.01.
104. Mine S. Ueda T. Hashimoto Y. Tanaka Y. Imoto T., High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements., FEBS Letters, 10.1016/S0014-5793(99)00332-4, 448, 1, 33-37, 488, 1, 33-37, 1999.01.
105. Ito HO. So T. Hirata M. Koga T. Ueda T. Imoto T, Tolerogenic activity of polyethylene glycol-conjugated lysozyme distinct from that of the native counterpart, Immunology, 93, 2, 200-207, 93, ,200-207, 1998.03.
106. Hashimoto Y. Miki T. Mukae M. Ueda T. Imoto T, Construction of a yeast expression system with positive selection for gene insertion in the absence of a specific phenotype, Gene, 10.1016/S0378-1119(97)00621-5, 207, 2, 167-170, 207,  , 167-170, 1998.01.
107. Abe Y. Ueda T. Kawano K. Tanaka Y. Imoto T, Detection of a local interaction of hen lysozyme under highly denaturing conditions using chemically 13C-enriched methionine resonance, The Journal of Biochemistry, 123, 2, 313-317, 123, , 313-317, 1998.01.
108. Ueda T. Tsurumaru R. Imoto T, Kinetic measurement of the interaction between a lysozymeand its immobilized substrate analogue by means of surface plasmon resonanc, The Journal of Biological Chemistry, 124, 4, 712-716, 124, , 712-716, 1998.01.
109. Kawamura S. Abe Y. Ueda T. Masumoto K. Imoto T. Yamasaki N. Kimura M., Investigation of the structural basis for thermostability of DNA-binding protein HU from Bacillus stearothermophilus, Journal of Biological Chemistry, 10.1074/jbc.273.32.19982, 273, 32, 19982-19987, 273, , 19982-19987, 1998.01.
110. Maeda Y. Ueda T. Imoto T., Effective renaturation of Met-1 lysozyme expressed in Escherichia coli as inclusion bodies., Protein and Peptide Letters, 5, 2, 67-74, 5, , 67-74, 1998.01.