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.


Full Text : http://www.dtic.mil/get-tr-doc/pdf?AD=ADA344173


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
      ORGANIC CHEMISTRY


Distribution Statement : APPROVED FOR PUBLIC RELEASE