A single adenosine with a neutral pKa in the ribosomal peptidyl transferase center.
Biochemical and crystallographic evidence suggests that 23S ribosomal RNA (rRNA) is the catalyst of peptide bond formation. To explore the mechanism of this reaction, we screened for nucleotides in Escherichia coli 23S rRNA that may have a perturbed pKa (where Ka is the acid constant) based on the pH dependence of dimethylsulfate modification. A single universally conserved A (number 2451) within the central loop of domain V has a near neutral pKa of 7.6 +/- 0.2, which is about the same as that reported for the peptidyl transferase reaction. In vivo mutational analysis of this nucleotide indicates that it has an essential role in ribosomal function. These results are consistent with a mechanism wherein the nucleotide base of A2451 serves as a general acid base during peptide bond formation.
Department of Molecular Biophysics and Biochemistry, Yale University, 260 Whitney Avenue, New Haven, CT 06520-8114, USA.
G W
GW
Muth
L
L
Ortoleva-Donnelly
S A
SA
Strobel
D000241:Adenosine / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002384:Catalysis; D004121:Dimethyl Sulfoxide; D004926:Escherichia coli; D006860:Hydrogen Bonding; D008745:Methylation; D009154:Mutation; D010452:Peptide Biosynthesis; D010458:Peptidyl Transferases / Q000737:chemistry* / Q000378:metabolism*; D011522:Protons; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*; D014372:Tubercidin / Q000378:metabolism
D011522:Protons; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; C018258:3-deazaadenosine; D010458:Peptidyl Transferases; D000241:Adenosine; D014372:Tubercidin; D004121:Dimethyl Sulfoxide