||Kiyota Sakai, Fumiko Matsuzaki, Lisa Wise, Yu Sakai, Sadanari Jindou, Hirofumi Ichinose, Naoki Takaya, Masashi Kato, Hiroyuki Wariishi, Motoyuki Shimizu, Biochemical characterization of CYP505D6, a self-sufficient cytochrome P450 from the white-rot fungus Phanerochaete chrysosporium, Applied and Environmental Microbiology, 10.1128/AEM.01091-18, 84, 22, 2018.11, The activity of a self-sufficient cytochrome P450 enzyme, CYP505D6, from the lignin-degrading basidiomycete Phanerochaete chrysosporium was characterized. Recombinant CYP505D6 was produced in Escherichia coli and purified. In the presence of NADPH, CYP505D6 used a series of saturated fatty alcohols with C9-18 carbon chain lengths as the substrates. Hydroxylation occurred at the ω-1 to ω-6 positions of such substrates with C9-15 carbon chain lengths, except for 1-dodecanol, which was hydroxylated at the ω-1 to ω-7 positions. Fatty acids were also substrates of CYP505D6. Based on the sequence alignment, the corresponding amino acid of Tyr51, which is located at the entrance to the active-site pocket in CYP102A1, was Val51 in CYP505D6. To understand the diverse hydroxylation mechanism, wild-type CYP505D6 and its V51Y variant and wild-type CYP102A1 and its Y51V variant were generated, and the products of their reaction with dodecanoic acid were analyzed. Compared with wild-type CYP505D6, its V51Y variant generated few products hydroxylated at the ω-4 to ω-6 positions. The products generated by wild-type CYP102A1 were hydroxylated at the ω-1 to ω-4 positions, whereas its Y51V variant generated ω-1 to ω-7 hydroxydodecanoic acids. These observations indicated that Val51 plays an important role in determining the regiospecificity of fatty acid hydroxylation, at least that at the ω-4 to ω-6 positions. Aromatic compounds, such as naphthalene and 1-naphthol, were also hydroxylated by CYP505D6. These findings highlight a unique broad substrate spectrum of CYP505D6, rendering it an attractive candidate enzyme for the biotechnological industry..
||Hiroyuki Kubota, Shinsuke Uda, Fumiko Matsuzaki, Yukiyo Yamauchi, Shinya Kuroda, In Vivo Decoding Mechanisms of the Temporal Patterns of Blood Insulin by the Insulin-AKT Pathway in the Liver, Cell Systems, 10.1016/j.cels.2018.05.013, 2018.07, Cells respond to various extracellular stimuli through a limited number of signaling pathways. One strategy to process such stimuli is to code the information into the temporal patterns of molecules. Although we showed that insulin selectively regulated molecules depending on its temporal patterns using Fao cells, the in vivo mechanism remains unknown. Here, we show how the insulin-AKT pathway processes the information encoded into the temporal patterns of blood insulin. We performed hyperinsulinemic-euglycemic clamp experiments and found that, in the liver, all temporal patterns of insulin are encoded into the insulin receptor, and downstream molecules selectively decode them through AKT. S6K selectively decodes the additional secretion information. G6Pase interprets the basal secretion information through FoxO1, while GSK3β decodes all secretion pattern information. Mathematical modeling revealed the mechanism via differences in network structures and from sensitivity and time constants. Given that almost all hormones exhibit distinct temporal patterns, temporal coding may be a general principle of system homeostasis by hormones. Kubota et al. show that the insulin-AKT pathway in the liver processes the information encoded into the temporal patterns of blood insulin and selectively regulates downstream molecules. Given that almost all hormones exhibit distinct temporal patterns, our study demonstrates the possibility of temporal coding as a general principle of systemic homeostasis by hormones..
||Masaki Matsumoto, Fumiko Matsuzaki, Kiyotaka Oshikawa, Naoki Goshima, Masatoshi Mori, Yoshifumi Kawamura, Koji Ogawa, Eriko Fukuda, Hirokazu Nakatsumi, Tohru Natsume, Kazuhiko Fukui, Katsuhisa Horimoto, Takeshi Nagashima, Ryo Funayama, Keiko Nakayama, Keiichi Nakayama, A large-scale targeted proteomics assay resource based on an in vitro human proteome, NATURE METHODS, 10.1038/nmeth.4116, 14, 3, 251-258, 2017.03.
||Yutaka Hashimoto, Michiko Shirane, Fumiko Matsuzaki, Shotaro Saita, Takafumi Ohnishi, Keiichi Nakayama, Protrudin Regulates Endoplasmic Reticulum Morphology and Function Associated with the Pathogenesis of Hereditary Spastic Paraplegia, JOURNAL OF BIOLOGICAL CHEMISTRY, 10.1074/jbc.M113.528687, 289, 19, 12946-12961, 2014.05.
||Fumiko Matsuzaki, Michiko Shirane, Masaki Matsumoto, Keiichi Nakayama, Protrudin serves as an adaptor molecule that connects KIF5 and its cargoes in vesicular transport during process formation, MOLECULAR BIOLOGY OF THE CELL, 10.1091/mbc.E11-01-0068, 22, 23, 4602-4620, 2011.12.
||Fumiko Matsuzaki, Motoyuki Shimizu, Hiroyuki Wariishi, Proteomic and metabolomic analyses of the white-rot fungus Phanerochaete chrysosporium exposed to exogenous benzoic acid, JOURNAL OF PROTEOME RESEARCH, 10.1021/pr700617s, 7, 6, 2342-2350, 2008.06.
||Fumiko Matsuzaki, Hiroyuki Wariishi, Molecular characterization of cytochrome P450 catalyzing hydroxylation of benzoates from the white-rot fungus Phaherochaete chrysosporzum, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 10.1016/j.bbrc.2005.07.013, 334, 4, 1184-1190, 2005.09.
||Fumiko Matsuzaki, Hiroyuki Wariishi, Functional diversity of cytochrome P450s of the white-rot fungus Phanerochaete chrysosporium, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 10.1016/j.bbrc.2004.09.062, 324, 1, 387-393, 2004.11.