Identification of the N‐tosyl‐L‐phenylalanyl chloromethylketone modification site in Thermus thermophilus elongation factor Tu
Open Access
- 6 November 1989
- journal article
- Published by Wiley in FEBS Letters
- Vol. 257 (2) , 219-222
- https://doi.org/10.1016/0014-5793(89)81538-8
Abstract
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.Keywords
This publication has 19 references indexed in Scilit:
- Modification of Bacillus subtilis elongation factor Tu by N‐tosyl‐L phenylalanyl chloromethane abolishes its ability to interact with the 3′‐terminal polynucleotide structure but not with the acyl bond in aminoacyl‐tRNAFEBS Letters, 1989
- Crystallization and preliminary X‐ray diffraction studies of intact EF‐Tu from Thermus aquaticus YT‐1FEBS Letters, 1988
- Sequence and identification of the nucleotide binding site for the elongation factor Tu fromThermus thermophilusHB8Nucleic Acids Research, 1987
- Structural homology between elongation factors EF - Tu from Bacillus stearothermophilus and Escherichia coli in the binding site for aminoacyl-tRNAEuropean Journal of Biochemistry, 1986
- The Elongation Factor EF-Tu and Its Two Encoding GenesProgress in Nucleic Acid Research and Molecular Biology, 1983
- Properties of Native and Nicked Elongation Factor Tu from Thermus thermophilus HB 8European Journal of Biochemistry, 1981
- A simplified procedure for the isolation of bacterial polypeptide elongation factor EF-TuAnalytical Biochemistry, 1980
- The binding site for the 3′‐terminus of aminoacyl‐tRNA in the molecule of elongation factor Tu from Escherichia coliFEBS Letters, 1979
- Studies on Polypeptide‐Chain‐Elongation Factors from an Extreme Thermophile, Thermus thermophilus HB8European Journal of Biochemistry, 1978
- Tosylphenylalanyl chloromethane-inhibitor of complex of S1S3-factors in cell-free protein-synthetizing system fromBacillus stearothermophilusFEBS Letters, 1971