Photoaffinity labeling of the primary fibrin polymerization site: isolation and characterization of a labeled cyanogen bromide fragment corresponding to gamma-chain residues 337-379.
Human fibrinogen and the plasmin-generated fibrinogen fragment D were photoaffinity labeled specifically with the peptide [14C]Gly-Pro-Arg-N(4-azido-2-nitrophenyl)Lys amide. In the case of fibrinogen, greater than 85% of the incorporated radioactivity was found in the gamma chain. Similarly, when fragment D (Mr, 90,000) was labeled with the same derivatized peptide, virtually all the radioactivity was found in the gamma-chain portion. The labeled fragment D was treated with CNBr and an initial purification was achieved by two gel-filtration steps. The labeled material was purified further by HPLC and was also compared with CNBr digests of unlabeled material. Amino acid analysis and gas-phase sequencing showed the labeled fragment to be gamma-chain residues 337-379.
Shimizu
A
A
Department of Chemistry, University of California, San Diego, La Jolla 92093-0634.
Nagel
G M
GM
Doolittle
R F
RF
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003488:Cyanogen Bromide / Q000737:chemistry; D005337:Fibrin / Q000737:chemistry*; D005340:Fibrinogen / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010777:Photochemistry; D011108:Polymers
D000596:Amino Acids; D010446:Peptide Fragments; D011108:Polymers; D005337:Fibrin; D005340:Fibrinogen; D003488:Cyanogen Bromide