A new strategy for primary structure determination of proteins: application to bovine beta-casein.
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.
Carles
C
C
Institut National de la Recherche Agronomique, CRJ, Jouy-en-Josas, France.
Huet
J C
JC
Ribadeau-Dumas
B
B
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
D002364:Caseins; D010446:Peptide Fragments; D010447:Peptide Hydrolases