A new nucleic acid-protein cross-linking reagent.

Molecular & general genetics : MGG ·Vol. 152 ·No. 3 ·1977-04-29 ·Pages 253-7

Oste C, Parfait R, Bollen A, Crichton RR

Abstract

A new photoactivable reagent is described, which allows the formation of RNA-protein cross-links via disulfide bridges in combination with mercaptobutyrimidate. The reconstituted L24 protein-23S RNA complex from the large subunit of E. coli ribosomes has been used as a model system for the cross-linking. The main advantages of the reagent are the absence of U.V. generated cross-links, since photoactivation is carried out at 360 nm, on one hand and the ease of cleavage of the cross-link by mild reduction (beta-mercaptoethanol) on the other.

MeSH Terms

Diazonium Compounds/pharmacology Indicators and Reagents Light Photochemistry Proteins RNA Sulfhydryl Compounds/pharmacology

Chemicals

Diazonium Compounds Indicators and Reagents Proteins Sulfhydryl Compounds RNA

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Oste C

Parfait R

Bollen A

Crichton R R

References (11)

11 references, click to expand

Purification of proteins from the 50S ribosomal subunit of Escherichia coli by ion-exchange chromatography.
Biochemistry. 1976 May 4;15(9):2007-17 PMID: 817738
Separation of large quantities of ribosomal subunits by zonal ultracentrifugation.
Eur J Biochem. 1970 Jan;12(1):113-6 PMID: 4907527
Structure and function of E. coli ribosomes. V. Reconstitution of functionally active 30S ribosomal particles from RNA and proteins.
Proc Natl Acad Sci U S A. 1968 Mar;59(3):777-84 PMID: 4868216
Methyl 4-mercaptobutyrimidate as a cleavable cross-linking reagent and its application to the Escherichia coli 30S ribosome.
Biochemistry. 1973 Aug 14;12(17):3266-73 PMID: 4581787
Ribosomal proteins. Isolation of proteins from 50S ribosomal subunits of Escherichia coli.
Eur J Biochem. 1971 Nov 11;23(1):12-6 PMID: 4942547
A native ribonucleoprotein complex from Escherichia coli ribosomes.
Proc Natl Acad Sci U S A. 1973 Mar;70(3):665-8 PMID: 4577133
Activities of B. stearothermophilus 50 S ribosomes reconstituted with prokaryotic and eukaryotic 5 S RNA.
FEBS Lett. 1973 Jul 15;33(3):315-9 PMID: 4580763
Photoaffinity labeling of 23 S rRNA in Escherichia coli ribosomes with poly(U)-coded ethyl 2-diazomalonyl-Phe-tRNA.
FEBS Lett. 1973 Dec 1;37(2):291-4 PMID: 4587218
Specific cross-linking of proteins S7 and L4 to ribosomal RNA, by UV irradiation of Escherichia coli ribosomal subunits.
Mol Gen Genet. 1975 Dec 9;141(4):343-55 PMID: 814400
Reductive methylation: a method for preparing functionally active radioactive ribosomes.
FEBS Lett. 1973 Nov 15;37(1):74-8 PMID: 4585034
Photo-induced joining of a transfer RNA with its cognate aminoacyl-transfer RNA synthetase.
J Mol Biol. 1974 Apr 25;84(4):503-13 PMID: 4840999

Article Info

Journal

Molecular & general genetics : MGG

Abbr.

Mol Gen Genet

ISSN

0026-8925

DOI

10.1007/BF00693078

Published

1977-04-29

Pages

253-7

Language

English

Region

Germany

NLM ID

0125036

Subset

External Links

PubMed source DOI: 10.1007/BF00693078

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