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Leucine-rich Repeats 2—4 (Leu60—Glu128) of Platelet Glycoprotein Ibα Regulate Shear-dependent Cell Adhesion to von Willebrand Factor.
- Source :
-
Journal of Biological Chemistry . 9/1/2006, Vol. 281 Issue 35, p26419-26423. 5p. 4 Graphs. - Publication Year :
- 2006
-
Abstract
- Glycoprotein (GP) Ib-IX-V binds von Willebrand factor (VWF), initiating thrombosis at high shear stress. The VWF-A1 domain binds the N-terminal domain of GPIbα (His¹-Glu282); this region contains seven leucine-rich repeats (LRR) plus N- and C-terminal flanking sequences and an anionic sequence containing three sulfated tyrosines. Our previous analysis of canine/human and human/canine chimeras of GPlba expressed on Chinese hamster ovary (CHO) cells demonstrated that LRR2-4(Leu60-Glu128) were crucial for GPIbα-dependent adhesion to VWF. Paradoxically, co-crystal structures of the GPIbα N-terminal domain and GPIbα-binding VWF-A1 under static conditions revealed that the LRR2-4 sequence made minimal contact with VWF-A1. To resolve the specific functional role of LRR2-4, we compared wild-type human GPIbα with human GPIbα containing a homology domain swap of canine for human sequence within Leu60-Glu128 and a reverse swap (canine GPIbα with human Leu60-Glu128) for the ability to support adhesion to VWF under flow. Binding of conformation-specific anti-GPIbα antibodies and VWF binding in the presence of botrocetin (which does not discriminate between species) confirmed equivalent expression of wild-type and mutant receptors in a functional form competent to bind ligand. Compared with CHO cells expressing wild-type GPIbα, cells expressing GPIbα, where human Leu60-Glu128 sequence was replaced by canine sequence, supported adhesion to VWF at low shear rates but became increasingly ineffective as shear increased from 50 to 2000 s-1. Together, these data demonstrate that LRR2-4, encompassing a pronounced negative charge patch on human GPIbα, is essential for GPIbα·VWF-dependent adhesion as hydrodynamic shear increases. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00219258
- Volume :
- 281
- Issue :
- 35
- Database :
- Academic Search Index
- Journal :
- Journal of Biological Chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 23134522
- Full Text :
- https://doi.org/10.1074/jbc.M604296200