|
|
Blood, 15 March 2008, Vol. 111, No. 6, pp. 3042-3049.
Prepublished online as a Blood First Edition Paper on November 1, 2007; DOI 10.1182/blood-2007-06-095042.
 Previous Article | Table of Contents | Next Article 
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
N-linked glycosylation of VWF modulates its interaction with ADAMTS13
Thomas A. J. McKinnon1,
Alain C. K. Chion1,
Alexander J. Millington1,
David A. Lane1, and
Mike A. Laffan1
1 Haematology Department, Imperial College of London, Hammersmith Hospital Campus, London, United Kingdom
We examined the role of N-linked glycan structures of VWF on its interaction with ADAMTS13. PNGase F digestion followed by lectin analysis demonstrated that more than 90% of VWF N-linked glycan chains could be removed from the molecule (PNG-VWF) without disruption of its multimeric structure or its ability to bind to collagen. PNG-VWF had an approximately 4-fold increased affinity for ADAMTS13 compared with control VWF. PNG-VWF was cleaved by ADAMTS13 faster than control VWF and was also proteolysed in the absence of urea. Occupancy of the N-linked glycan sites at N1515 and N1574 and their presentation of ABO(H) blood group sugars were confirmed with an isolated tryptic fragment. Recombinant VWF was mutated to prevent glycosylation at these sites. Mutation of N1515 did not alter ADAMTS13 binding or increase rate of ADAMTS13 proteolysis. Mutation of N1574 increased the susceptibility of VWF to ADAMTS13 proteolysis and allowed cleavage in the absence of urea. Mutation of N1574 in the isolated recombinant VWF-A2 domain also increased binding and ADAMTS13 proteolysis. These data demonstrate that the N-linked glycans of VWF have a modulatory effect on the interaction with ADAMTS13. At least part of this effect is conformational, but steric hindrance may also be important.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
A. F. Riddell, K. Gomez, C. M. Millar, G. Mellars, S. Gill, S. A. Brown, M. Sutherland, M. A. Laffan, and T. A. J. McKinnon
Characterization of W1745C and S1783A: 2 novel mutations causing defective collagen binding in the A3 domain of von Willebrand factor
Blood,
October 15, 2009;
114(16):
3489 - 3496.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Zanardelli, A. C. K. Chion, E. Groot, P. J. Lenting, T. A. J. McKinnon, M. A. Laffan, M. Tseng, and D. A. Lane
A novel binding site for ADAMTS13 constitutively exposed on the surface of globular VWF
Blood,
September 24, 2009;
114(13):
2819 - 2828.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. de Groot, A. Bardhan, N. Ramroop, D. A. Lane, and J. T. B. Crawley
Essential role of the disintegrin-like domain in ADAMTS13 function
Blood,
May 28, 2009;
113(22):
5609 - 5616.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. D. Gardner, C. K. N. K. Chion, R. de Groot, A. Shah, J. T. B. Crawley, and D. A. Lane
A functional calcium-binding site in the metalloprotease domain of ADAMTS13
Blood,
January 29, 2009;
113(5):
1149 - 1157.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Zhou and H.-M. Tsai
N-Glycans of ADAMTS13 modulate its secretion and von Willebrand factor cleaving activity
Blood,
January 22, 2009;
113(4):
929 - 935.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
