Identification of the N‐tosyl‐L‐phenylalanyl chloromethylketone modification site in Thermus thermophilus elongation factor Tu

FEBS Letters

Peter, Marcus E.²  Jonák, Jiří¹  Brockmöller, Jürgen³  Sprinzl, Mathias² 

[1] Institute of Molecular Genetics, Czechoslovak Acad. Sci., Flemingovo nam. 2,16637 Praha 6, Czechoslovakia;Laboratorium für Biochemie, Universität Bayreuth, Postfach 101251, D-8580 Bayreuth, FRG;Max-Planck-Institutfür Molekulare Genetik, Ihnestrasse 63, D-1000 Berlin 33, Germany

关键词: GTP-binding protein;    Protein biosynthesis;    Elongation factor Tu;    N-Tosyl-L-phenylalanyl chloromethylketone;    (Thermus thermophilus);    EF-Tu and EF-Ts;    elongation factors Tu and Ts;    aatRNA;    aminoacyl-tRNA;    TPCK;    Mtosyl-L-phenylalanyl chloromethylketone;    PTH;    phenylthiohydantoin;   

DOI  :  10.1016/0014-5793(89)81538-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

EF-Tu from Thermus thermophilus was first labelled with N-[¹⁴C]tosyl-L-phenylalaninechloromethylketone and then cleaved by the combined action of CNBr and trypsin. The resulting peptides were separated by reversed-phase HPLC. Analysis of the isolated, labelled peptide led to the identification of a sequence which was identical to residues 76–88 in T. thermophilus EF-Tu. The TPCK reactive site is at Cys-82. Kinetic measurements f the incorporation of TPCK into native EF-Tu and EF-Tu nicked at position Arg-59 were performed. The results provide evidence that the cleavage of the peptide bond between Arg-59 and Gly-60 does not lead to a dramatic conformational change of EF-Tu at the aa-tRNA binding site.

【 授权许可】

Unknown