Substrate Recognition Mechanism of Carboxypeptidase Y(Biochemistry & Molecular Biology)
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概要
- 論文の詳細を見る
To clarify the substrate-recognition mechanism of carboxypeptidase Y, Fmoc-(Glu)_nAla-OH (n=1 to 6), Fmoc-(Glu)_nAla-NH_2 (1 to 5), and Fmoc-Lys(Glu)_3Ala-NH_2 were synthesized, and kinetic parameters for these substrates were measured. K_m for Fmoc-peptides significantly decreased as peptide length increased from n = 1 to n = 5 with only slight changes in k_<cat>. K_m for Fmoc-(Glu)_<5,6>Ala-OH were almost the same as one for protein substrates described previously (Nakase et al., Bull. Chem. Soc. Jpn., 73, 2587-2590). These results show that the enzyme has six subsites (S_1' and S_1-S_5). Each subsite affinity calculated from the K_m revealed subsite properties, and from the differences of subsite affinity between pH 6.5 and 5.0, the residues in each subsite were predicted. For Fmoc-peptide amide substrates, the priorities of amidase and carboxamide peptidase activities were dependent on the substrate. It is likely that the interactions between side chains of peptide and subsites compensate for the lack of P_1'-S_1' interaction, so the amidase activity prevailed for Fmoc-(Glu)_<3,5>Ala-NH_2. These results suggest that these subsites contribute extensively to substrate recognition rather than a hydrogen bond network.
- 社団法人日本農芸化学会の論文
- 2001-11-23
著者
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NAKASE Hiroshi
Division of Gastroenterology, Nishi-Kobe Medical Center
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Nakase Hiroshi
Departments Of Gastroenterology And Endoscopic Medicine Kyoto University Hospital
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Nakase Hiroshi
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Nakase Hiroshi
Department Of Gastroenterology & Hepatology Graduate School Of Medicine Kyoto University
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UENO Hiroshi
Division of Pathology, Central Clinical Laboratory
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HAYASHI Rikimaru
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
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Ueno H
Laboratory Of Biomacromolecular Chemistry Division Of Applied Life Sciences The Graduate School Of A
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Nakase Hiroshi
Department Of Gastroenterology And Hepatology Kyoto University Graduate School Of Medicine
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Murata Shoichi
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Hayashi R
Laboratory Of Biomacromolecular Chemistry Division Of Applied Life Sciences The Graduate School Of A
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Hayashi Rikimaru
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Hayashi R
Kyoto Univ. Kyoto
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Ueno Hiroshi
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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