Studies on the measurement of protein S in plasma

Bovill, Edwin G.; Landesman, M; Busch, Shirley; Fregeau, G R; Mann, KG; Tracy, Russ

doi:10.1093/clinchem/37.10.1708

Cited by 17 publications

(10 citation statements)

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“…An early study using gel‐filtered bovine plasma suggested that there were three forms of PS in addition to PS–C4BP complexes [27]. In another study, most of the free PS in human plasma had an apparent mass of dimers during gel filtration, although conformational considerations could apply [28]. These are in partial agreement with our finding that free plasma PS consists of monomers and multimers.…”

Section: Discussionsupporting

confidence: 89%

Plasma contains protein S monomers and multimers with similar direct anticoagulant activity

Heeb

Radtke²,

Fernández

et al. 2006

Journal of Thrombosis and Haemostasis

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Summary. Background: Protein S (PS) has activated protein C-independent, direct anticoagulant activity (PS-direct). We reported that both multimers and monomers of affinity-purified PS have PS-direct similar to that in plasma, in contrast to another report. Objective: We extended our studies to establish the molecular forms and activity of plasma PS. Methods: Novel ELISAs were developed that could detect only multimeric, not monomeric, PS because they employed the same monoclonal antibody for capture and detection. PS forms were also examined on native PAGE immunoblots. A new activity assay for PS-direct was applied to plasma and gel-filtered plasma fractions. Results: Plasma PS multimers were clearly demonstrated using the ELISAs; 30-60% of free plasma PS appeared to be multimeric, a proportion similar to that of affinity-purified PS. On immunoblots, plasma PS multimers were more easily detected after gel filtration; plasma PS monomers and several apparent multimers comigrated with respective forms of affinity-purified PS. Antigen elution profiles after gel filtration of plasma revealed at least one major peak of apparent PS multimers (40-55% of free PS appeared multimeric). Biotinfactor Xa could bind to both plasma PS monomers and multimers. Strong plasma PS-direct was demonstrated, and plasma PS monomers, multimers, and PS-C4b-binding protein complexes each reconstituted PS-depleted plasma to similar levels of PS-direct. Conclusion: Our data are in disagreement with a report that monomeric purified PS has little PS-direct and that only monomeric PS exists in plasma. We find that both affinity-purified and plasma PS exist as monomers and multimers with similar PS-direct.

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Section: Discussionsupporting

confidence: 89%

Plasma contains protein S monomers and multimers with similar direct anticoagulant activity

Heeb

Radtke²,

Fernández

et al. 2006

Journal of Thrombosis and Haemostasis

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“…All assays were performed in the investigators' laboratories either by ELISA: FV antigen [coefficient of variation (CV) 5.8%][21], FVIII antigen (CV 7.8%), FIX antigen (CV 10%), VWF (CV 3%) [22], protein Z (CV 6.5%) [24], ZPI (CV 7.2%) [24], APC–α1AT complex (CV 12.4%) [25] and APC–PCI complex (CV 11.7%) [25], or radioimmunoassay: prothrombin (CV 8%) [26], antithrombin (CV 5%) [27,28], protein S (CV 9.8%) [29], F1.2 (CV 8%) [28], PCP (CV 14%) [28] and FPA (CV 8%) [28], the Clauss method for fibrinogen (CV 1.7%) [30,31] using the ST4 instrument (Diagnostica Stago, Parsipanny, NJ, USA), a clot‐based functional assay for protein C: (CV 5.5%) [20,23] or micro latex bead agglutination for d ‐dimer (CV 9.2%) (Biomerieux, Durham, NC, USA) [32,33].…”

Section: Methodsmentioning

confidence: 99%

Heritability of plasma concentrations of clotting factors and measures of a prethrombotic state in a protein C‐deficient family

Vossen

Hasstedt

Rosendaal

et al. 2004

Journal of Thrombosis and Haemostasis

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of plasma concentrations of clotting factors and measures of a prethrombotic state in a protein C-de®cient family. J Thromb Haemost 2004; 2: 242±7.Summary. Background: Earlier studies found strong support for a genetic basis for regulation of coagulation factor levels and measures of a prethrombotic state (D-dimer, prothrombin fragment 1.2). Objectives: Estimation of how much of the variation in the levels of coagulation factors and measures of a prethrombotic state, including measures of protein C activation and inactivation, could be attributed to heritability and household effect. Patients and methods: Blood samples were collected from 330 members of a large kindred of French-Canadian origin with type I protein C de®ciency. Heritability and common household effect were estimated for plasma concentrations of prothrombin, factor (F)V, factor VIII, factor (F)IX, ®brinogen, von Willebrand factor (VWF), antithrombin, protein C, protein S, protein Z, protein Z-dependent protease inhibitor (ZPI), ®brinopeptide A (FPA), protein C activation peptide (PCP), activated protein C±protein C inhibitor complex (APC±PCI), activated protein C±a 1 -antitrypsin complex (APC±a1AT), prothrombin fragment 1.2 (F1.2) and D-dimer, using the variance component method in sequential oligo-genic linkage analysis routines (SOLAR). Results: The highest heritability was found for measures of thrombin activity (PCP and FPA). High estimates were also found for prothrombin, FV, FIX, protein C, protein Z, ZPI, APC±PCI and APC±a1AT. An important in¯uence of shared household effect on phenotypic variation was found for VWF, antithrombin, protein S and F1.2. Conclusions: We found strong evidence for the heritability of single coagulation factors and measures of a prethrombotic state. Hemostatic markers with statistically signi®cant heritability constitute potential targets for the identi®cation of novel genes involved in the control of quantitative trait loci.

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“…The assays for factor VIIc (CV = 5.3%), factor VIIag (CV = 6.0%), factor VIIIc (CV = 9.7%), factor IXc (CV = 5.8%), factor Xc (4.7%), fibrinogen (CV = 2.9%), tissue factor pathway inhibitor (TFPI; CV = 9.5%), antithrombin-III (AT-III; CV = 4.5%), protein C antigen (CV = 2.5%), total protein S (9.9%), free protein S (CV = 6.7%), factor XIa-␣ 1 -anti-trypsin complex (FXIa-␣ 1 -AT; CV = 6.1%), C-reactive protein (CRP; CV = 8.9%), prothrombin fragment 1-2 (F1-2; CV = 8.5%), fibrinopeptide A (FPA; CV = 8.4%), fibrin fragment D-dimer (CV = 7.0%), ␣ 1 acid glycoprotein (CV = 7.0%), plasminogen (CV = 3.6%), tissue plasminogen activator (tPA; CV = 7.0%), plasminogen activator inhibitor-1 (PAI-1; CV = 8.4%), plasmin-␣ 2 -antiplasmin (PAP; CV = 3.0%), and tPA/PAI complex (CV = 14.3%) have been previously described (4,30,(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47).…”

Section: Antigen Levelsmentioning

confidence: 95%

No Association of Plasma Prothrombin Concentration or the G20210A Mutation with Incident Cardiovascular Disease

Smiles

Jenny²,

Tang³

et al. 2002

Thromb Haemost

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SummaryProthrombin is a key factor in blood clotting, a process intimately involved in thrombotic disease. We assessed prothrombin levels and G20210A genotype in a case-control study within the Cardiovascular Health Study. Cases included angina, myocardial infarction, stroke, and the presence of MRI-detectable infarcts (n ≈ 250 each). Populationbased controls free of clinical cardiovascular disease (CVD) (n ≈ 500) and a subset free of clinical and subclinical CVD (n ≈ 250) were used for comparison. The 20210 A allele, frequency 2.9%, was associated with higher mean prothrombin levels: 166.3 vs. 139.5 µg/ml (P <0.001). Significant correlates of prothrombin included gender, plasma lipids, other vitamin K-dependent proteins, and inflammatory markers, but not race, smoking, hypertension, diabetes, measures of subclinical CVD, or markers of procoagulant activity. Compared to controls, neither genotype nor prothrombin level was associated with any CVD case group. We conclude that, in the elderly, neither prothrombin level nor 20210 genotype were associated with either CVD risk factors or events. This is consistent with the lack of association of prothrombin levels with measures of underlying CVD or procoagulant markers.

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Studies on the measurement of protein S in plasma

Cited by 17 publications

References 0 publications

Plasma contains protein S monomers and multimers with similar direct anticoagulant activity

Plasma contains protein S monomers and multimers with similar direct anticoagulant activity

Heritability of plasma concentrations of clotting factors and measures of a prethrombotic state in a protein C‐deficient family

No Association of Plasma Prothrombin Concentration or the G20210A Mutation with Incident Cardiovascular Disease

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