Phylogenetic and Functional Analysis of Histidine Residues Essential for pH-dependent Multimerization of von Willebrand Factor

Dang, Luke T.; Purvis, Angie R.; Huang, Ren Huai; Westfield, Lisa A.; Sadler, J. Evan

doi:10.1074/jbc.m111.249151

Cited by 27 publications

(47 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pH-dependent control of VWF multimerization in the N-terminal VWD1-D2-D′D3 domains depends on two conserved histidine residues of the VWD2 domain, His 395 and His 460 , respectively (12). Although the histidine residue His 395 is present in the VWD2 domain of MUC2, the second histidine residue His 460 is absent and may influence the different oligomeric state of these two structurally related proteins.…”

Section: Discussionmentioning

confidence: 99%

“…3D reconstructions based on transmission electron microscopy (TEM) images showed that the N-terminal VWF helix consists of 4.2 repeating units per turn (11). This pH-dependent packing required protonation of conserved histidine residues (12). Because the packing of VWF tubules depends solely on the N-terminal part of VWF, it was suggested that the rest of the VWF extended radially from the helix axis (11).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin

Ambort

Johansson

Gustafsson

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

285

396

View full text Add to dashboard Cite

MUC2, the major colonic mucin, forms large polymers by N-terminal trimerization and C-terminal dimerization. Although the assembly process for MUC2 is established, it is not known how MUC2 is packed in the regulated secretory granulae of the goblet cell. When the N-terminal VWD1-D2-D′D3 domains (MUC2-N) were expressed in a goblet-like cell line, the protein was stored together with fulllength MUC2. By mimicking the pH and calcium conditions of the secretory pathway we analyzed purified MUC2-N by gel filtration, density gradient centrifugation, and transmission electron microscopy. At pH 7.4 the MUC2-N trimer eluted as a single peak by gel filtration. At pH 6.2 with Ca 2+ it formed large aggregates that did not enter the gel filtration column but were made visible after density gradient centrifugation. Electron microscopy studies revealed that the aggregates were composed of rings also observed in secretory granulae of colon tissue sections. The MUC2-N aggregates were dissolved by removing Ca 2+ and raising pH. After release from goblet cells, the unfolded full-length MUC2 formed stratified layers. These findings suggest a model for mucin packing in the granulae and the mechanism for mucin release, unfolding, and expansion.ucins are large glycoproteins that coat the surface of cells in the respiratory, digestive, and urogenital tracts (1, 2). Their main function is protection of epithelial cells from infection and physical injury. Mucins are characterized by mucin domains that are heavily O-glycosylated on the protein sequence rich in proline, threonine, and serine, therefore called PTS domains (3). These domains have little interspecies sequence conservation but often have tandemly repeated amino acid sequences that vary in number and length (3). There are several mucin types; the gelforming mucins are the only ones that form large polymers. In humans there are four gel-forming mucin genes that are known to be expressed, MUC2 in the intestine (4), MUC5AC in lungs and stomach, MUC5B in lungs and saliva, and MUC6 in stomach (1).MUC2 mucin is the major component of the mucus (mixture of mucins and other associated proteins) in the small and large intestine (2). In colon this is organized into two layers: an inner, densely packed layer that is attached to the epithelium that is impermeable to bacteria, and an outer, easily removable loose layer that is the habitat for the commensal bacteria (5). Human MUC2 mucin has 5,179 amino acids and contains multiple domains arranged in the following order (Fig. 1A): von Willebrand D1 domain (VWD1), VWD2, VWD′D3, (VWD1-D2-D′D3), first CysD, small PTS, second CysD, large PTS (tandemly repeated), C-terminal VWD4 followed by VWB, VWC, and a cystine-knot domain (CK) (4). The primary translational product of full-length MUC2 is quickly dimerized in the endoplasmic reticulum (ER) via disulfide bonds in the CK domain (6). The dimers pass into the Golgi apparatus, where the two PTS domains become O-glycosylated to form the two mucin domains. In the trans-Golgi network the glycosylated ...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin

Ambort

Johansson

Gustafsson

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

285

396

View full text Add to dashboard Cite

show abstract

“…More recent evidence suggests that protonation of one or more histidine residues in VWFpp mediates the pH dependence of multimerization. 31 By using histidine to alanine mutagenesis, Dang et al demonstrated that replacement of histidine 395 or 460 in VWFpp caused a profound defect in inter-subunit disulfide formation, suggesting that these 2 residues in VWFpp contribute to the pH-dependent regulation of VWF multimer assembly. 31 VWFpp clearly has a critical role in the multimerization of VWF.…”

Section: Life Cycle Of Vwf and Vwfppmentioning

confidence: 99%

von Willebrand factor propeptide: biology and clinical utility

Haberichter

2015

Blood

View full text Add to dashboard Cite

von Willebrand factor (VWF) is a large multimeric glycoprotein that mediates the attachment of platelets to damaged endothelium and also serves as the carrier protein for coagulation factor VIII (FVIII), protecting it from proteolytic degradation. Quantitative or qualitative defects in VWF result in von Willebrand disease (VWD), a common inherited bleeding disorder. VWF is synthesized with a very large propeptide (VWFpp) that is critical for intracellular processing of VWF. VWFpp actively participates in the process of VWF multimerization and is essential for trafficking of VWF to the regulated storage pathway. Mutations identified within VWFpp in VWD patients are associated with altered VWF structure and function. The assay of plasma VWFpp has clinical utility in assessing acute and chronic vascular perturbation associated with diseases such as thrombotic thrombocytopenic purpura, sepsis, and diabetes among others. VWFpp assay also has clear utility in the diagnosis of VWD subtypes, particularly in discriminating true type 3 subjects from type 1C (reduced plasma survival of VWF), which is clinically important and has implications for therapeutic treatment.

show abstract

“…Enabled by the lower pH of the Golgi apparatus, this intrinsic oxidoreductase function is activated by protonation of histidine residues adjacent to these CGLC sequences. 6 Mutations within these sequences inhibit multimerization but do not affect dimerization. Therefore, dimerization does not appear to be performed by an intrinsic oxidoreductase activity within VWF.…”

Section: Introductionmentioning

confidence: 99%

von Willebrand factor is dimerized by protein disulfide isomerase

Lippok

Kolšek

Löf

et al. 2016

Blood

View full text Add to dashboard Cite

Key Points• The protein disulfide isomerase is involved in VWF dimerization by initiating disulfide bond formation at cysteines 2771 and 2773.• von Willebrand diseaseassociated mutations in the dimerization domain of von Willebrand factor disturb processing by the protein disulfide isomerase.Multimeric von Willebrand factor (VWF) is essential for primary hemostasis. The biosynthesis of VWF high-molecular-weight multimers requires spatial separation of each step because of varying pH value requirements. VWF is dimerized in the endoplasmic reticulum by formation of disulfide bonds between the C-terminal cysteine knot (CK) domains of 2 monomers. Here, we investigated the basic question of which protein catalyzes the dimerization. We examined the putative interaction of VWF and the protein disulfide isomerase PDIA1, which has previously been used to visualize endoplasmic reticulum localization of VWF. Excitingly, we were able to visualize the PDI-VWF dimer complex by high-resolution stochastic optical reconstruction microscopy and atomic force microscopy. We proved and quantified direct binding of PDIA1 to VWF, using microscale thermophoresis and fluorescence correlation spectroscopy (dissociation constants K D 5 236 6 66 nM and K D 5 282 6 123 nM by microscale thermophoresis and fluorescence correlation spectroscopy, respectively). The similar K D (258 6 104 nM) measured for PDI interaction with the isolated CK domain and the atomic force microscopy images strongly indicate that PDIA1 binds exclusively to the CK domain, suggesting a key role of PDIA1 in VWF dimerization. On the basis of protein-protein docking and molecular dynamics simulations, combined with fluorescence microscopy studies of VWF CK-domain mutants, we suggest the following mechanism of VWF dimerization: PDI initiates VWF dimerization by forming the first 2 disulfide bonds Cys2771-27739 and Cys27719-2773. Subsequently, the third bond, Cys2811-28119, is formed, presumably to protect the first 2 bonds from reduction, thereby rendering dimerization irreversible. This study deepens our understanding of the mechanism of VWF dimerization and the pathophysiological consequences of its inhibition. (Blood.

show abstract

Phylogenetic and Functional Analysis of Histidine Residues Essential for pH-dependent Multimerization of von Willebrand Factor

Cited by 27 publications

References 38 publications

Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin

Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin

von Willebrand factor propeptide: biology and clinical utility

von Willebrand factor is dimerized by protein disulfide isomerase

Contact Info

Product

Resources

About