title: Mass spectrometry based analysis of human plasma-derived factor X revealed novel post-translational modifications.
abstract: Human coagulation factor X is a central component of the blood coagulation cascade that converts, under its activated form, prothrombin into thrombin. Generation of thrombin is the final step of the clotting cascade that leads to the clot by polymerization of fibrinogen molecules into a fibrin network. Today, research of new by-passing agents of the coagulation may contribute to an increased interest for human factor X, which may, in consequence, lead to the need of a more exhaustive picture of its structural features. Several post-translational modifications of human factor X such as γ-carboxylation/β-hydroxylation of the N-terminal light chain and N-/O-glycosylation of the activation peptide have been described. But, so far as we know, no comprehensive studies of its post-translational modifications have been reported. In this article we report an exhaustive structural analysis of human factor X by mass spectrometry using successive protein and peptide mapping. Surprisingly, human factor X was found to be mostly O-glucosylated on its light chain at Ser106 position, Ser9 of its activation peptide is phosphorylated at about 30% and its C-terminal heavy chain is fully O-glycosylated at Thr249 by a mucin-type O-glycan (HexNAc-Hex-NeuAc). The knowledge of these post-translational modifications is mandatory for the development of recombinant molecules.
authors.affiliation: Analytical Department of LFB Biotechnologies, 3 Avenue Des Tropiques, 91942, Courtaboeuf Cedex, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Chevreux
authors.affiliation: Analytical Department of LFB Biotechnologies, 3 Avenue Des Tropiques, 91942, Courtaboeuf Cedex, France.
authors.forename: Nolwenn
authors.initials: N
authors.lastname: Tilly
authors.affiliation: Analytical Department of LFB Biotechnologies, 3 Avenue Des Tropiques, 91942, Courtaboeuf Cedex, France.
authors.forename: Valegh
authors.initials: V
authors.lastname: Faid
authors.affiliation: Analytical Department of LFB Biotechnologies, 3 Avenue Des Tropiques, 91942, Courtaboeuf Cedex, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bihoreau
mesh_terms: D005170:Factor X / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational
chemical_list: D010455:Peptides; D005170:Factor X
keywords: O-glucosylation; O-glycosylation; coagulation factor X; mass spectrometry; phosphorylation