Cross-linking site in fibrinogen for alpha 2-plasmin inhibitor.
A plasma proteinase inhibitor, alpha 2-plasmin inhibitor (alpha 2PI), is cross-linked with alpha chain of fibrin(ogen) by activated coagulation Factor XIII (plasma transglutaminase). alpha 2PI serves only as a glutamine substrate (amine acceptor) for activated Factor XIII in the cross-linking reaction, and the cross-linking occurs between Gln-2 of the alpha 2PI molecule and a lysine residue (amine donor) of fibrin(ogen) alpha chain, whose position was investigated. alpha 2PI and fibrinogen were reacted by activated Factor XIII. The resulting alpha 2PI fibrinogen A alpha chain complex was separated and subjected to two cycles of Edman degradation using phenyl isothiocyanate for the first cycle and dimethylaminoazobenzene-isothiocyanate for the second cycle. The aqueous phase after the cleavage stage of the second cycle, containing dimethylaminoazobenzene-thiohydantoin-Gln cross-linked with A alpha chain, was subjected to CNBr fragmentation and tryptic digestion. Only one of the peptides was found to have the peak of absorbance at 420 nm, indicating the presence of dimethylaminoazobenzene-thiohydantoin-Gln in that peptide. The peptide was identified as corresponding to residues Asn-290-Arg-348 of A alpha chain by analyses of the NH2-terminal amino acid sequence and amino acid composition. The peptide contains a single lysine at position 303, indicating that Lys-303 of fibrinogen A alpha chain is the lysine residue that forms a cross-link with Gln-2 of alpha 2PI.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D003488:Cyanogen Bromide; D005176:Factor XIII / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis / Q000494:pharmacology; D011503:Transglutaminases; D014357:Trypsin; D000979:alpha-2-Antiplasmin / Q000378:metabolism*
D000596:Amino Acids; D010446:Peptide Fragments; D000979:alpha-2-Antiplasmin; D005340:Fibrinogen; D005176:Factor XIII; D011503:Transglutaminases; D014357:Trypsin; D003488:Cyanogen Bromide