Reductive Alkylation of Ribosomes As a Probe to the Topography of Ribosomal Proteins

Overview

Journal Biochem J

Specialty Biochemistry

Date 1974 Dec 1

PMID 4462744

Citations 4

Authors

G Moore

R R Crichton

Affiliations

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Abstract

Escherichia coli ribosomes were treated with a number of different aldehydes of various sizes in the presence of NaBH(4). After incorporation of either (3)H or (14)C, the ribosomal proteins were separated by two-dimensional polyacrylamide-gel electrophoresis and the extent of alkylation of the lysine residues in each protein was measured. The same pattern of alkylation was observed with the four reagents used, namely formaldehyde, acetone, benzaldehyde and 3,4,5-trimethoxybenzaldehyde. Every protein in 30S and 50S subunits was modified, although there was considerable variation in the degree of alkylation of individual proteins. A topographical classification of ribosomal proteins is presented, based on the degree of exposure of lysine residues. The data indicate that every protein of the ribosome has at least one lysine residue exposed at or near the surface of the ribonucleo-protein complex.

Citing Articles

Radioactive labelling of ribosomal proteins with reductive alkylation and its use in studying ribosome-cytosol interactions.

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New low resolution model for 50S subunit of Escherichia coli ribosomes.

Stuhrmann H, Haas J, Ibel K, Wolf B, Koch M, Parfait R Proc Natl Acad Sci U S A. 1976; 73(7):2379-83.

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Identification of components of the streptomycin-binding center of E. coli MRE 600 ribosomes by photo-affinity labelling.

Girshovich A, Bochkareva E, Ovchinnikov Y Mol Gen Genet. 1976; 144(2):205-12.

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Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.

Towbin H, Staehelin T, Gordon J Proc Natl Acad Sci U S A. 1979; 76(9):4350-4.

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