Crosslinking of elongation factor Tu to tRNAPhe by trans‐diamminedichloroplatinum (II) Characterization of two crosslinking sites on EF‐Tu

Abstract

In a preceding paper [(1987) Nucleic Acids Res. 15, 5787–5801], we have used trans-diamminedichloroplatinum (II) to induce reversible RNA-protein crosslinks within the ternary EF-Tu/GTP/Phe-tRNA^Phe complex and have identified two crosslinking sites on the tRNA. The aim of the present paper is to determine the crosslinking sites on EF-Tu. Two tryptic peptides located in domain I could be identified, a major one (residues 45–74) and a minor one (residues 117–154). The use of Staphylococcus aureus V8 protease led to the isolation of two major peptides (residues 56–68 and 64–68) and one minor peptide (118–124). These results are discussed in the light of the current knowledge of the topography of the EF-Tu/tRNA complex.