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

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 fe...

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Published inThrombosis 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
LanguageEnglish
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
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Summary: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.
ISSN:0340-6245
DOI:10.1160/TH06-03-0138