pKa of adenine 2451 in the ribosomal peptidyl transferase center remains elusive

RNA. 2001 Oct;7(10):1365-9.

Abstract

The universally conserved A2451 of 23S rRNA has been proposed to participate directly in the catalysis of peptide bond formation in the ribosomal peptidyl transferase center. An unusually high, near neutral, pKa of A2451 is a prerequisite for its action as a general acid-base catalyst. Increased reactivity of A2451 to dimethylsulfate (DMS) at pH 8.5 compared to pH 6.5 was taken as evidence that the pKa of this nucleotide falls within this pH range. Structural data suggested that the interaction between A2451 and G2447 in the ribosome is responsible for A2451 pKa perturbation. In contrast to expectation, our studies did not show pH dependence of A2451 dimethylsulfate modification in ribosomes of Thermus aquaticus and Mycobacterium smegmatis. Other rRNA regions, however, showed major alterations in DMS reactivity at pH 8.5 compared to pH 6.5, suggesting that conformational rearrangements in the structure of the large ribosomal subunit may occur upon the pH shift. The G2447U mutant of M. smegmatis was viable, indicating that the G2447-A2451 interaction is not critical for the ribosome function. We concluded that the proposed unusual pKa of A2451, if existing, may not be crucial for the ribosome activity and that the previously reported pH-dependent alterations in the DMS modification of A2451 do not necessarily reveal an unusual pKa of this nucleotide.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenine / metabolism*
  • Base Sequence
  • Molecular Sequence Data
  • Mycobacterium smegmatis / enzymology
  • Nucleic Acid Conformation
  • Peptidyl Transferases / chemistry
  • Peptidyl Transferases / metabolism*
  • RNA, Bacterial / chemistry
  • RNA, Ribosomal, 23S / chemistry
  • Ribosomes / enzymology*
  • Thermus / enzymology

Substances

  • RNA, Bacterial
  • RNA, Ribosomal, 23S
  • Peptidyl Transferases
  • Adenine