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Identification of neighbouring proteins in the 30 S ribosomes of E. coli
Author(s) -
Barritault D.,
Expert-Bezançon A.,
Milet M.,
Háyes D.H.
Publication year - 1975
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(75)80469-8
Subject(s) - marie curie , curie , humanities , curie temperature , physics , philosophy , condensed matter physics , ferromagnetism , european union , business , economic policy
Bifunctional reagents have been widely used recently in studies of neighbourhood relations between proteins of E. coli ribosomes [l-9] . The composition of most of the cross-linked protein complexes so far observed after treatment of 30 S ribosomes with these reagents are consistent with proximity relationships implicit in the 30 S assembly map [lo] , with the protein compositions of ribonucleoprotein fragments isolated after ribonuclease digestion of 30 S ribosomes [ 1 l-1 31 and with protein neighbourhoods deduced from analyses of functional sites in 30 S ribosomes [ 141. The possibility that cross-linked complexes might be formed in some cases as a consequence of structural deformation of ribosomes in the presence of bifunctional reagents seems unlikely because it has been shown that functionally active 30 S subunits can be reconstituted in vitro from 16 S rRNA and protein mixtures in which 30 S proteins S 5 and S 8 are replaced by a dimethyl adipimidate cross-linked complexofS5andS8 [15]. Two major difficulties are encountered in the use of bifunctional reagents for the study of ribosome structure; isolation of pure complexes from the mixture of reaction products and the identification of the proteins present in isolated complexes. Here we describe briefly experimental techniques which overcome these difficulties and their application to the study of E. coli 30 S ribosome subunits.