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PMID: 1093183 Published · ppublish English Journal Article

Affinity labeling of the ribosomal decoding site with an AUG-substrate analog.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 72 ·No. 4 ·1975-04-00 ·Pages 1505-9

Pongs O, Lanka E

Abstract

The trinucleotide AUG was condensed at the 5'-end with N-bromoacetyl-p-aminophenylphosphate. This bromoactylated AUG analog reacted irreversibly with the mRNA binding site of Escherichia coli 70S ribosomes. After reaction of 70S ribosomes with the AUG analog, labeled 30S subunits could be isolated that were programmed for initiation-factor-dependent binding of fMet-tRNAfMet. This shows that this AUG-affinity label reacted in the decoding site for fMet-tRNAfMet. By combination of sodium dodecyl sulfate-, Sarkosyl-, and ureapolyacrylamide gel electrophoresis the AUG-affinity label was found to be irreversibly bound to ribosomal proteins S4, the ram gene product, and S18.

MeSH Terms
Binding Sites Centrifugation, Density Gradient Electrophoresis, Polyacrylamide Gel Escherichia coli/metabolism Kinetics N-Formylmethionine/metabolism Oligonucleotides/metabolism Peptide Initiation Factors RNA, Bacterial/metabolism RNA, Messenger/metabolism RNA, Transfer/metabolism Receptors, Drug Ribosomes/metabolism Structure-Activity Relationship
Chemicals
Oligonucleotides Peptide Initiation Factors RNA, Bacterial RNA, Messenger Receptors, Drug N-Formylmethionine RNA, Transfer
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Pongs O
Lanka E
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21 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.72.4.1505
Published
1975-04-00
Pages
1505-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC432565
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.72.4.1505 PMC Full Text View journal details
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