The peptidyltransferase centre of the Escherichia coli ribosome. The histidine of protein L16 affects the reconstitution and control of the active centre but is not essential for release-factor-mediated peptidyl-tRNA hydrolysis and peptide bond formation.
Modification of the Escherichia coli 50S ribosomal subunit with histidine-specific diethyl pyrocarbonate affects peptide bond formation and release-factor-dependent peptidyl-tRNA hydrolysis. Unmodified L16 can restore activity to a split protein fraction from the altered subunit but other proteins of the core also contain histidine residues important for the activity of the peptidyltransferase centre. When isolated and purified by centrifugation, particles reconstituted with unmodified proteins and modified L16 do not retain the altered L16. The modified protein does mediate the partial restoration of peptide bond formation and release-factor-2 activities to these particles. It must be exerting its effect during the assembly of the peptidyltransferase centre in the reconstituted particle. A particle could be reconstituted which lacks L16 and has significant activity in peptide bond formation and peptidyl-tRNA hydrolysis. L16 stimulates these activities. A tighter ribosomal binding of the release factor 2, dependent upon the absence of protein L11, can in part compensate for the loss of activity of the peptidyltransferase centre when it is assembled with either modified L16 or in the absence of L16. The protein and its histidine residue seem important, therefore, for the peptidyltransferase centre to be formed in the correct conformation but not essential for activity once the centre is assembled.
Tate
W P
WP
Sumpter
V G
VG
Trotman
C N
CN
Herold
M
M
Nierhaus
K H
KH
D000217:Acyltransferases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D006639:Histidine / Q000502:physiology; D006868:Hydrolysis; D010454:Peptide Termination Factors / Q000502:physiology; D010455:Peptides / Q000378:metabolism; D010458:Peptidyl Transferases / Q000378:metabolism*; D011691:Puromycin / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000201:enzymology*
D001426:Bacterial Proteins; D010454:Peptide Termination Factors; D010455:Peptides; D012346:RNA, Transfer, Amino Acyl; D012269:Ribosomal Proteins; C041460:peptide chain termination release factor 2; C027937:ribosomal protein L16; C019037:tRNA, peptidyl-; D011691:Puromycin; D006639:Histidine; D000217:Acyltransferases; D010458:Peptidyl Transferases