Molecular characterization of the prokaryotic efp gene product involved in a peptidyltransferase reaction

被引:28
作者
Aoki, H
Adams, SL
Turner, MA
Ganoza, MC
机构
[1] UNIV TORONTO, BANTING & BEST DEPT MED RES, TORONTO, ON M5G 1L6, CANADA
[2] HOSP SICK CHILDREN, DIV BIOCHEM RES, TORONTO, ON M5G 1X8, CANADA
关键词
ribosomes; translation; peptidyltransferase; antibiotics;
D O I
10.1016/S0300-9084(97)87619-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The translation factor EF-P is required for efficient prokaryotic peptide bond synthesis on 70S ribosomes from fMet-tRNA(Met)(f). This protein has been purified from Escherichia coli cells and the gene, efp, encoding it has been cloned and sequenced, We have isolated recombinant clones which overexpress a protein that co-migrates with purified EF-P upon SDS-PAGE analysis. Using these-clones, we report the purification, crystallization and initial characterization of the efp gene product. The mechanism by which EF-P stimulates peptide-bond synthesis was studied using several antibiotics that inhibit translocation, peptide-bond synthesis and decoding. The stimulation of peptidyltransferase by EF-P was not inhibited by antibiotics that affect translocation and occupation of the A site (in the elongation state), ie thiostrepton, viomycin, neomycin and fusidic acid but was inhibited by streptomycin as well as by inhibitors of peptidyltransferase, chloramphenicol and lincomycin. This observation and the requirement for L16 but not for the L7/L12 nor L6 or L11 r-proteins suggest that the binding site for EF-P may overlap the peptidyltransferase center of the ribosome.
引用
收藏
页码:7 / 11
页数:5
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