title: Two missense mutations identified in venous thrombosis patients impair the inhibitory function of the protein Z dependent protease inhibitor.
abstract: Protein Z-dependent protease inhibitor (ZPI) is a plasma inhibitor of factor (F)Xa and FXIa. In an earlier study, five mutations were identified within the ZPI gene of venous thrombosis patients and healthy controls. Two of these were nonsense mutations and three were missense mutations in important regions of the protein. Here we report that two of these latter three mutations, F145L and Q384R, impair the inhibitory function of ZPI in vitro. Recombinant wild-type and mutant proteins were prepared; stability in response to thermal challenge was similar. Inhibition of FXa in the presence of the cofactor protein Z was reduced 68-fold by the Q384R mutant; inhibition of FXIa by the F145L mutant was reduced two- to three-fold compared to the wild-type ZPI. An analysis of all five ZPI mutations was undertaken in a cohort of venous thrombosis patients (n=550) compared to healthy controls (n=600). Overall, there was a modest increase in incidence of these mutations in the thrombosis group (odds ratio 2.0, 1.05-3.7, p=0.044). However, in contrast to W324X (nonsense mutation), the Q384R missense mutation and R88X nonsense mutation were evenly distributed in patients and controls; F145L was rare. The final mutation (S143Y) was also rare and did not significantly alter ZPI function in laboratory studies. The F145L and particularly the Q384R mutation impaired the function of the coagulation inhibitor ZPI; however, there was no convincing association between these mutations and venous thrombosis risk. The functional role for ZPI in vivo has yet to be clarified.
authors.affiliation: Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand. l.young@auckland.ac.nz
authors.forename: Laura K
authors.initials: LK
authors.lastname: Young
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Birch
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Browett
authors.forename: Paul B
authors.initials: PB
authors.lastname: Coughlin
authors.forename: Anita J
authors.initials: AJ
authors.lastname: Horvath
authors.forename: Neil S
authors.initials: NS
authors.lastname: Van de Water
authors.forename: Paul A
authors.initials: PA
authors.lastname: Ockelford
authors.forename: Paul L
authors.initials: PL
authors.lastname: Harper
mesh_terms: D000328:Adult; D001777:Blood Coagulation / Q000235:genetics*; D016022:Case-Control Studies; D016009:Chi-Square Distribution; D004795:Enzyme Stability; D015945:Factor XIa / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D009520:New Zealand; D016017:Odds Ratio; D010641:Phenotype; D011487:Protein Conformation; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000378:metabolism; D018570:Risk Assessment; D012307:Risk Factors; D015843:Serpins / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D013329:Structure-Activity Relationship; D020246:Venous Thrombosis / Q000097:blood / Q000201:enzymology / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; C416370:SERPINA10 protein, human; D015843:Serpins; D015945:Factor XIa; D015951:Factor Xa