title: A proteomic analysis of changes in prothrombin and plasma proteins associated with the G20210A mutation.
abstract: The G-->A mutation at position 20210 of the prothrombin gene, localized in the 3'-polyadenylation untranslated region of the mRNA, is a recognized genetic risk factor for venous thromboembolism. The mechanism by which this base change confers an increased risk of thrombosis compared to noncarriers is undefined. Studies on the mRNA suggest enhanced cleavage site recognition and a change in the location of the 3'-cleavage/polyadenylation reaction, but no defined model has been proposed. The present study, based on proteomic investigation by two-dimensional gel electrophoresis and electrospray ionization (ESI) tandem mass spectrometry (MS/MS) protein identification, suggests that the G20210A mutation is associated with increased glycosylation of prothrombin, which confers greater stability to the protein. Additionally, proteomic investigation of pooled plasma showed that expression levels of six spots, three of them identified by ESI MS/MS, were altered in subjects carrying the mutation, suggesting a possible cooperative effect in the thrombotic risk increment induced by the mutation.
authors.affiliation: Institute of Molecular Bioimaging and Physiology, CNR, Via F.lli Cervi 93, 20090 Segrate, Milan, Italy. cecilia.gelfi@ibfm.cnr.it
authors.forename: Cecilia
authors.initials: C
authors.lastname: Gelfi
authors.forename: Agnese
authors.initials: A
authors.lastname: Viganò
authors.forename: Marilena
authors.initials: M
authors.lastname: Ripamonti
authors.forename: Robin
authors.initials: R
authors.lastname: Wait
authors.forename: Shajna
authors.initials: S
authors.lastname: Begum
authors.forename: Eugenia
authors.initials: E
authors.lastname: Biguzzi
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Castaman
authors.forename: Elena Maria
authors.initials: EM
authors.lastname: Faioni
mesh_terms: D001798:Blood Proteins / Q000737:chemistry* / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006031:Glycosylation; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D009154:Mutation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D011516:Prothrombin / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D012307:Risk Factors; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001798:Blood Proteins; D011506:Proteins; D012333:RNA, Messenger; D011516:Prothrombin