title: Effects of introducing fibrinogen Aalpha character into the factor XIII activation peptide segment.
abstract: The formation of a blood clot involves the interplay of thrombin, fibrinogen, and Factor XIII. Thrombin cleaves fibrinopeptides A and B from the N-termini of the fibrinogen Aalpha and Bbeta chains. Fibrin monomers are generated that then polymerize into a noncovalently associated network. By hydrolyzing the Factor XIII activation peptide segment at the R37-G38 peptide bond, thrombin assists in activating the transglutaminase FXIIIa that incorporates cross-links into the fibrin clot. In this work, the kinetic effects of introducing fibrinogen Aalpha character into the FXIII AP segment were examined. Approximately 25% of fibrinogen Aalpha is phosphorylated at Ser3, producing a segment with improved binding to thrombin. FXIII AP ((22)AEDDL(26)) has sequence properties in common with Fbg Aalpha ((1)ADSpGE(5)). Kinetic benefits to FXIII AP cleavage were explored by extending FXIII AP (28-41) to FXIII AP (22-41) and examining peptides with D24, D24S, D24Sp, and D24Sp P27G. These modifications did not provide the same kinetic advantages that were observed with Fbg Aalpha (1-20) S3p. Such results further emphasize that FXIII AP derives most of its substrate specificity from the P(9)-P(1) segment. To enhance the kinetic properties of FXIII AP (28-41), we introduced substitutions at the P(9), P(4), and P(3) positions. Studies reveal that FXIII AP (28-41) V29F, V34G, V35G exhibits kinetic improvements that are comparable to those of FXIII AP V29F, V34L and approach those of Fbg Aalpha (7-20). Selective changes to the FXIII AP segment sequence may be used to design FXIII species that can be activated more or less readily.
authors.affiliation: Department of Chemistry, University of Louisville, 2320 South Brook Street, Louisville, Kentucky 40292, USA.
authors.forename: Madhavi A
authors.initials: MA
authors.lastname: Jadhav
authors.forename: Giulia
authors.initials: G
authors.lastname: Isetti
authors.forename: Toni A
authors.initials: TA
authors.lastname: Trumbo
authors.forename: Muriel C
authors.initials: MC
authors.lastname: Maurer
mesh_terms: D004789:Enzyme Activation; D005176:Factor XIII / Q000235:genetics / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments; D013379:Substrate Specificity; D013917:Thrombin / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism
chemical_list: D010446:Peptide Fragments; C464794:fibrinogen Aalpha; D005340:Fibrinogen; D005176:Factor XIII; D011503:Transglutaminases; D013917:Thrombin