Identification of a new truncated form and deamidation products of fibrinopeptide B released by thrombin from human fibrinogen

• Published in: Thrombosis and haemostasis Vol. 96; no. 3; p. 302
• Main Authors: Cardinali, Barbara, Damonte, Gianluca, Melone, Luca, Salis, Annalisa, Tosetti, Francesca, Rocco, Mattia, Profumo, Aldo
• Format: Journal Article
• Language: English
• Published: Germany 01.09.2006
• Subjects: Blood Coagulation Factors - chemistry; Chemistry, Physical - methods; Chromatography; Chromatography, High Pressure Liquid; Electrophoresis, Capillary; Fibrinogen - chemistry; Fibrinogen - metabolism; Fibrinopeptide B - chemistry; Humans; Mass Spectrometry; Spectrometry, Mass, Electrospray Ionization; Thrombin - chemistry
• ISSN: 0340-6245
• DOI: 10.1160/TH06-03-0138

Quantification of fibrinopeptides release is widely used to investigate fibrinogen activation, and standard chromatographic or capillary electrophoretic procedures are readily available. However, in the analyses of fibrinopeptide mixtures derived from the action of thrombin on human fibrinogen, a few unidentified peaks are usually present. The composition of these peaks was studied by reverse-phase HPLC/MS, revealing a single major anomalous peptide having a molecular mass of 1384.4. A further MS/MS analysis allowed the identification of this form, as a Nterminally truncated fibrinopeptide B (fpB) lacking the first two residues (pyroglutamic acid and glycine). This previously unidentified, relatively low-abundance form ( approximately 7%) has been found consistently in our fibrinopeptides preparations, and analysis of the parent Bbeta-chain suggest that it is likely present in circulating fibrinogen. In addition, deamidated forms of all fpB species (including desArgB), resulting from the conversion of asparagine to aspartic acid, were also identified. Overall, these previously unreported forms constitute a substantial amount of fpB (up to approximately 17% of the total), and should be taken into account for a reliable quantitative analysis of fpB release.