Accession Number : ADA344173
Title : Cleavage Mapping the Topology of Protein Folding Intermediates
Descriptive Note : Final rept.
Corporate Author : YALE UNIV NEW HAVEN CT
Personal Author(s) : Ledman, David W.
Report Date : 13 MAR 1995
Pagination or Media Count : 52
Abstract : In order to identify regions of the polypeptide chain that are brought close together by the protein fold, our laboratory devised a scheme in which, in the presence of reductant, reactive oxygen species are generated at an EDTA-Fe moiety covalently attached to unique cysteine residues in a series of single cysteine mutants of the protein under study. Only surface residues that do not participate in side chain interactions are mutated. The mutants undergo a thiol exchange reaction in the presence of EPD-Fe that results in the attachment of the EDTA-Fe moiety. The free radicals generated at the iron center promote intramolecular, conformation dependent cleavage of the peptide backbone.
Descriptors : *PEPTIDES , *PROTEINS , CRYSTAL STRUCTURE , EXCHANGE REACTIONS , INTERACTIONS , ENZYMES , MUTATIONS , TOPOLOGY , MAPPING , MOLECULAR PROPERTIES , FOLDING , CYSTEINE , AMINO ACIDS , STAPHYLOCOCCUS , CLEAVAGE , THIOLS , NUCLEASE.
Subject Categories : BIOCHEMISRTY
Distribution Statement : APPROVED FOR PUBLIC RELEASE