Abstract
The method of affinity labeling has been used to identify protein components of 50S ribosomal subunits involved in peptidyl transferase activity. E. coli 50S ribosomal subunits were mapped by reaction with the N-bromoacetyl analog of chloramphenicol, an antibiotic known to interact specifically with the active center of the enzyme. The synthetic analog competes with chloramphenicol in binding to 50S ribosomal subunits and inhibits peptidyl transferase activity. It attaches covalently to the ribosome under appropriate conditions and causes an irreversible loss in peptidyl transferase activity. The reagent specifically alkylates cysteine residues of proteins L2 and L27.
MeSH Terms
Acetamides/metabolism
Acyltransferases/antagonists & inhibitors,metabolism
Alkylation
Anti-Bacterial Agents/metabolism
Bacterial Proteins/analysis,biosynthesis
Binding, Competitive
Bromine
Carbon Isotopes
Chloramphenicol/metabolism
Cysteine/metabolism
Escherichia coli/enzymology
Ethylmaleimide/metabolism
Methionine/metabolism
Methods
Peptide Elongation Factors/antagonists & inhibitors,metabolism
Peptides
Phenethylamines/metabolism
Phenylalanine/metabolism
Ribosomes/enzymology
Succinimides/metabolism
Chemicals
Acetamides
Anti-Bacterial Agents
Bacterial Proteins
Carbon Isotopes
Peptide Elongation Factors
Peptides
Phenethylamines
Succinimides
Phenylalanine
Chloramphenicol
Methionine
Acyltransferases
Cysteine
Ethylmaleimide
Bromine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sonenberg N
Wilchek M
Zamir A
References (19)
19 references, click to expand
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