Crosslinking of elongation factor Tu to tRNA(Phe) by trans-diamminedichloroplatinum (II). Characterization of two crosslinking sites on EF-Tu.
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.
Metz-Boutigue
M H
MH
Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Reinbolt
J
J
Ebel
J P
JP
Ehresmann
C
C
Ehresmann
B
B
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002713:Chlorine; D002845:Chromatography; D002945:Cisplatin / Q000378:metabolism*; D003432:Cross-Linking Reagents; D004226:Dithioerythritol / Q000494:pharmacology; D004926:Escherichia coli / Q000032:analysis; D005056:Euglena gracilis / Q000032:analysis; D008969:Molecular Sequence Data; D010444:Peptide Elongation Factor Tu / Q000378:metabolism*; D010446:Peptide Fragments; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases; D013824:Thermus / Q000032:analysis; D014357:Trypsin
D000596:Amino Acids; D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D012345:RNA, Transfer, Amino Acid-Specific; D012360:RNA, Transfer, Phe; C034697:transplatin; D002713:Chlorine; D004226:Dithioerythritol; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D010444:Peptide Elongation Factor Tu; D002945:Cisplatin