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
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| 刊名 | JOURNAL OF BIOLOGICAL CHEMISTRY
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| 出版日期 | 2004-05-14 |
| 卷号 | 279期号:20页码:21128-21134 |
| ISSN号 | 0021-9258 |
| DOI | 10.1074/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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