Abstract：
:Aminopeptidase B (APB, EC 3.4.11.6) preferentially hydrolyzes the N-terminal basic amino acids of synthetic and peptide substrates and requires a physiological concentration of NaCl for optimal activity. In this study, we used site-directed mutagenesis and molecular modeling to search for an amino acid residue that is critical for the enzymatic properties of human APB. Substitution of Phe297 with Tyr caused a significant decrease in hydrolytic activity toward synthetic and peptide substrates as well as chloride anion sensitivity. Molecular modeling suggests that Phe297 contributes to the construction of the substrate pocket of APB, which is wide enough to hold a chloride anion and allow the interaction of Gln169 with the N-terminal Arg residue of the substrate through bridging with the chloride anion. These results indicate that Phe297 is crucial for the optimal enzymatic activity and chloride anion sensitivity of APB via formation of the optimal structure of the catalytic pocket.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Ohnishi A,Watanabe J,Ogawa Y,Goto Y,Hattori A,Tsujimoto Mdoi
10.1021/acs.biochem.5b00964subject
Has Abstractpub_date
2015-10-06 00:00:00pages
6062-70issue
39eissn
0006-2960issn
1520-4995journal_volume
54pub_type
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