title: A new strategy for primary structure determination of proteins: application to bovine beta-casein.
abstract: A new approach has been developed for sequencing proteins. A radioactive label is attached specifically to the C-terminus of the protein. The labelled molecule is subjected to varying proteolysis conditions. From the electrophoretic patterns (SDS-PAGE) of the hydrolysates, appropriate cleavage conditions are selected, giving labelled peptides of different lengths which are purified. The labelled peptides are sequenced in order of increasing size (from 1 to n), peptide (i) being sequenced until the N-terminal sequence of peptide (i-1) is encountered. This approach allows the determination of a complete protein sequence with a minimal number of Edman cycles. The method was successfully applied to bovine beta-casein (209 residues) which was completely resequenced with only 239 Edman cycles.
authors.affiliation: Institut National de la Recherche Agronomique, CRJ, Jouy-en-Josas, France.
authors.forename: C
authors.initials: C
authors.lastname: Carles
authors.forename: J C
authors.initials: JC
authors.lastname: Huet
authors.forename: B
authors.initials: B
authors.lastname: Ribadeau-Dumas
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002364:Caseins; D002417:Cattle; D008722:Methods; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010447:Peptide Hydrolases
chemical_list: D002364:Caseins; D010446:Peptide Fragments; D010447:Peptide Hydrolases