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