Control of Von Willebrand Factor Multimer Size by Thrombospondin-1

Xie, Lijuan; Chesterman, Colin N.; Hogg, Philip J.

doi:10.1084/jem.193.12.1341

Cited by 124 publications

(105 citation statements)

References 40 publications

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“…In the extracellular space, the Ser-700 polymorphism may alter interactions of the C-terminal region of TSP-1 with hemostatic proteins such as thrombin (31) and von Willebrand factor (32,33), which bind covalently to TSP-1 via disulfide bonding at the C terminus. Recently, the presence of the Ser-700 polymorphism in TSP-1 has been shown to increase binding of TSP-1 to fibrinogen on the platelet surface, resulting in increased platelet aggregation (30).…”

Section: The Tr1 Constructs Contain An Intact Tb 3ϩ -Binding Site-mentioning

confidence: 99%

A Polymorphism in Thrombospondin-1 Associated with Familial Premature Coronary Artery Disease Alters Ca2+ Binding

Hannah

Misenheimer²,

Pranghofer³

et al. 2004

Journal of Biological Chemistry

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A single nucleotide polymorphism that results in substitution at residue 700 of a serine (Ser-700) for an asparagine (Asn-700) in thrombospondin-1 is associated with familial premature coronary artery disease. The polymorphism is located in the first of 13 Ca 2؉ -binding motifs, within a consensus sequence in which Asn-700 likely coordinates Ca 2؉ . Equilibrium dialysis of constructs comprised of the adjoining epidermal growth factor-like module and the Ca 2؉ -binding region (E3Ca) demonstrated that E3Ca Ser-700 binds significantly less Ca 2؉ than E3Ca Asn-700 at low [Ca 2؉ ]. The hypothesis that this difference is due to loss of a binding site in Ser-700 protein was tested with truncations of E3Ca containing four (Tr4), three (Tr3), two (Tr2), or one (Tr1) N-terminal Ca 2؉ -binding motifs. The Ser-700 truncation constructs bound 1 fewer Ca 2؉ than matching Asn-700 constructs and exhibited decreased binding affinities. Intrinsic fluorescence of a tryptophan at residue 698 (Trp-698) in the most N-terminal motif was cooperatively quenched by the addition of Ca 2؉ to Asn-700 Tr2, Tr3, and Tr4 constructs. In Ser-700 constructs, quenching of Trp-698 was incomplete in the Tr2 and Tr3 constructs and complete only in the Tr4 construct. Ca 2؉ -induced quenching of Ser-700 constructs required higher [Ca 2؉ ] and was slower as shown in stopped-flow experiments than quenching of Asn-700 constructs. Such differences were not found with Tb 3؉ , which quenched the fluorescence of Asn-700 and Ser-700 constructs equivalently. Thus, the Ser-700 polymorphism alters a rapidly filled, high affinity Ca 2؉ -binding site in the first Ca 2؉ -binding motif. Slower Ca 2؉ binding to adjoining motifs partly compensates for the change.

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Section: The Tr1 Constructs Contain An Intact Tb 3ϩ -Binding Site-mentioning

confidence: 99%

A Polymorphism in Thrombospondin-1 Associated with Familial Premature Coronary Artery Disease Alters Ca2+ Binding

Hannah

Misenheimer²,

Pranghofer³

et al. 2004

Journal of Biological Chemistry

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“…22 Equal concentrations (0.5 g/mL) of wild type (wt) or mutant VWF or normal pooled plasma diluted in PBS were incubated with 100 M malemide-PEO 2 -Biotin (MPB; Thermo) for 10 minutes at room temperature and the reaction quenched with 200 M GSH. Samples were then diluted with PBS-T to final concentration 0.25 g/mL and applied to MaxiSorp plates that had been previously coated with polyclonal anti-VWF antibodies and incubated for 1 hour at room temperature.…”

Section: Analysis Of Free Thiol Content Of Vwfmentioning

confidence: 99%

Specific N-linked glycosylation sites modulate synthesis and secretion of von Willebrand factor

McKinnon¹,

Goode²,

Birdsey

et al. 2010

Blood

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We examined the role that N-linked glycans play in the synthesis and expression of von Willebrand Factor (VWF). Blocking the addition of N-linked glycans (NLGs) or inhibiting initial glycan processing prevented secretion of VWF. To determine whether specific glycosylation sites were important, the 16 VWF N-linked glycosylation sites were mutated followed by expression in HEK293T cells. Four NLG mutants affected VWF expression: N99Q (D1 domain), N857Q (D' domain), N2400Q (B1 domain), and N2790Q (CK domain) either abolished or reduced secretion of VWF and this was confirmed by metabolic labeling. Multimer analysis of mutant N2790Q cell lysate revealed an increase in VWF monomers, which was also observed when the isolated CK domain was expressed with N2790 mutated. Immunofluorescence microscopy showed that mutants N99Q, N857Q, and N2790Q were primarily retained within the ER, producing only few pseudo Weibel-Palade bodies over longer time periods compared with wtVWF. All the variants also showed an increase in free thiol reactivity. This was greatest with N857Q and D4-C2 NLG mutants, which had approximately 6-fold and 3-to 4-fold more free thiol reactivity than wtVWF. These data provide further evidence of the critical role that individual N-linked glycans play in determining VWF synthesis and expression. (Blood. 2010;116(4):640-648) Introductionvon Willebrand factor (VWF) is a large multimeric plasma glycoprotein essential for normal hemostasis, acting firstly by supporting platelet adhesion to surfaces at sites of vascular injury and secondly as the carrier molecule for pro-coagulant Factor VIII (FVIII). 1,2 Synthesis of VWF is limited to megakaryocytes and endothelial cells. 3 The pre-pro-VWF molecule comprises a 22 amino acid signal peptide, a 741 amino acid propeptide, and a 2050 amino acid mature subunit. The pro-VWF monomer is composed of 4 types of domains (A-D) arranged as follows: NH 2 -D1-D2-D'-D3-A1-A2-A3-D4-B1-B2-B3-C1-C2-CK-COOH. 1 VWF multimers are formed by C-and N-terminal intermolecular disulphide bonds, 4,5 with the largest multimers exceeding 2 ϫ 10 4 kDa and having the greatest adhesive activity. 6 During synthesis, VWF undergoes extensive posttranslational modification resulting in the addition of 12 N-linked and 10 O-linked glycosylation sites per mature monomer. 7 Furthermore, the propeptide also contains 4 potential N-linked glycosylation sites although it is not known if these are used. In total, carbohydrate accounts for approximately 20% of the molecular weight of VWF. 8 N-linked glycosylation is one of the most common co/ posttranslational modifications. It is characterized by the addition of a carbohydrate moiety to the protein via a beta-glycosidic linkage between an N-acetylglucosamine residue and the ␦-amide of an asparagine residue in the sequence NXS/T (or in some cases NXC), 9 where X is any amino acid except proline. 10 This takes place after translocation into the endoplasmic reticulum (ER) when the enzyme oligosaccharyltransferase (OST) transfers a preformed 14-saccharide core...

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“…But at least a subset of these antibodies also shows features of autoreactivity. 2 Parallels between these 2 syndromes are well recognized, 3 but the final link between them has remained enigmatic. In this issue of Blood, Sikara et al demonstrate in a series of in vitro experiments an interaction between ␤2-GPI and PF4.…”

Section: Opposites Attract ------------------------------------------mentioning

confidence: 99%

“…Consistent with the notion, an early report has shown that thiol(s) in the VWF A3 domain is targeted by a reductase activity associated with thrombospondin-1. 3 The question remains as to whether different reactive oxygen species differentially modify specific amino acids within a VWF multimer, leading to different functional outcomes. Second, does oxidation also affect VWF adhesion activity?…”

Section: Jing-fei Dong Baylor College Of Medicinementioning

confidence: 99%

Oxidative stress on VWF proteolysis

Dong

2010

Blood

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Control of Von Willebrand Factor Multimer Size by Thrombospondin-1

Cited by 124 publications

References 40 publications

A Polymorphism in Thrombospondin-1 Associated with Familial Premature Coronary Artery Disease Alters Ca2+ Binding

A Polymorphism in Thrombospondin-1 Associated with Familial Premature Coronary Artery Disease Alters Ca2+ Binding

Specific N-linked glycosylation sites modulate synthesis and secretion of von Willebrand factor

Oxidative stress on VWF proteolysis

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