中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Mutations at the S1 sites of methionine aminopeptidases from Escherichia coli and Homo sapiens reveal the residues critical for substrate specificity

文献类型:期刊论文

作者Li, JY; Cui, YM; Chen, LL; Gu, M; Li, J; Nan, FJ; Ye, QZ
刊名JOURNAL OF BIOLOGICAL CHEMISTRY
出版日期2004-05-14
卷号279期号:20页码:21128-21134
ISSN号0021-9258
DOI10.1074/jbc.M401679200
文献子类Article
英文摘要Methionine aminopeptidase (MetAP) catalyzes the removal of methionine from newly synthesized polypeptides. MetAP carries out this cleavage with high precision, and Met is the only natural amino acid residue at the N terminus that is accepted, although type I and type II MetAPs use two different sets of residues to form the hydrophobic S1 site. Characteristics of the S1 binding pocket in type I MetAP were investigated by systematic mutation of each of the seven S1 residues in Escherichia coli MetAP type I (EcMetAP1) and human MetAP type I (HsMetAP1). We found that Tyr-65 and Trp-221 in EcMetAP1, as well as the corresponding residues Phe-197 and Trp-352 in HsMetAP1, were essential for the hydrolysis of a thiopeptolide substrate, Met-S-Gly-Phe. Mutation of Phe-191 to Ala in HsMetAP1 caused inactivity in contrast to the full activity of EcMetAP1(Y62A), which may suggest a subtle difference between the two type I enzymes. The more striking finding is that mutation of Cys-70 in EcMetAP1 or Cys-202 in HsMetAP1 opens up the S1 pocket. The thiopeptolides Leu-S-Gly-Phe and Phe-S-Gly-Phe, with previously unacceptable Leu or Phe as the N-terminal residue, became efficient substrates of EcMetAP1(C70A) and HsMetAP1(C202A). The relaxed specificity shown in these S1 site mutants for the N-terminal residues was confirmed by hydrolysis of peptide substrates and inhibition by reaction products. The structural features at the enzyme active site will be useful information for designing specific MetAP inhibitors for therapeutic applications.
WOS关键词SACCHAROMYCES-CEREVISIAE ; PYROCOCCUS-FURIOSUS ; MOLECULAR-CLONING ; CRYSTAL-STRUCTURE ; INHIBITORS ; GENE ; FUMAGILLIN ; PROTEINS ; OVEREXPRESSION ; TRANSFERASE
资助项目NCRR NIH HHS[P20 RR 015563]
WOS研究方向Biochemistry & Molecular Biology
语种英语
WOS记录号WOS:000221273800070
出版者AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
源URL[http://119.78.100.183/handle/2S10ELR8/274077]  
专题国家新药筛选中心
通讯作者Nan, FJ
作者单位1.Chinese Acad Sci, Chinese Natl Ctr Drug Screening, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
2.Univ Kansas, Hight Throughput Screening Lab, Higuchi Biosci Ctr, Lawrence, KS 66047 USA
推荐引用方式
GB/T 7714
Li, JY,Cui, YM,Chen, LL,et al. Mutations at the S1 sites of methionine aminopeptidases from Escherichia coli and Homo sapiens reveal the residues critical for substrate specificity[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2004,279(20):21128-21134.
APA Li, JY.,Cui, YM.,Chen, LL.,Gu, M.,Li, J.,...&Ye, QZ.(2004).Mutations at the S1 sites of methionine aminopeptidases from Escherichia coli and Homo sapiens reveal the residues critical for substrate specificity.JOURNAL OF BIOLOGICAL CHEMISTRY,279(20),21128-21134.
MLA Li, JY,et al."Mutations at the S1 sites of methionine aminopeptidases from Escherichia coli and Homo sapiens reveal the residues critical for substrate specificity".JOURNAL OF BIOLOGICAL CHEMISTRY 279.20(2004):21128-21134.

入库方式: OAI收割

来源:上海药物研究所

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