Imperial College London
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Emeritus Professor David Lane

Faculty of Medicine, Department of Immunology and Inflammation

Emeritus Professor of Haematology
 
 
 
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Contact

 

+44 (0)20 3313 2295d.lane

 
 
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Location

 

Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{de:2015:10.1182/blood-2014-08-594556,
author = {de, Groot R and Lane, DA and Crawley, JTB},
doi = {10.1182/blood-2014-08-594556},
journal = {Blood},
pages = {1968--1975},
title = {The role of the ADAMTS13 cysteine-rich domain in VWF binding and proteolysis},
url = {http://dx.doi.org/10.1182/blood-2014-08-594556},
volume = {125},
year = {2015}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - ADAMTS13 proteolytically regulates the platelet-tethering function of von Willebrand factor (VWF). ADAMTS13 function is dependent upon multiple exosites that specifically bind the unraveled VWF A2 domain and enable proteolysis. We carried out a comprehensive functional analysis of the ADAMTS13 cysteine-rich (Cys-rich) domain using engineered glycans, sequence swaps, and single point mutations in this domain. Mutagenesis of Cys-rich domain–charged residues had no major effect on ADAMTS13 function, and 5 out of 6 engineered glycans on the Cys-rich domain also had no effect on ADAMTS13 function. However, a glycan attached at position 476 appreciably reduced both VWF binding and proteolysis. Substitution of Cys-rich sequences for the corresponding regions in ADAMTS1 identified a hydrophobic pocket involving residues Gly471-Val474 as being of critical importance for both VWF binding and proteolysis. Substitution of hydrophobic VWF A2 domain residues to serine in a region (residues 1642-1659) previously postulated to interact with the Cys-rich domain revealed the functional importance of VWF residues Ile1642, Trp1644, Ile1649, Leu1650, and Ile1651. Furthermore, the functional deficit of the ADAMTS13 Cys-rich Gly471-Val474 variant was dependent on these same hydrophobic VWF residues, suggesting that these regions form complementary binding sites that directly interact to enhance the efficiency of the proteolytic reaction.
AU  - de,Groot R
AU  - Lane,DA
AU  - Crawley,JTB
DO  - 10.1182/blood-2014-08-594556
EP  - 1975
PY  - 2015///
SN  - 0006-4971
SP  - 1968
TI  - The role of the ADAMTS13 cysteine-rich domain in VWF binding and proteolysis
T2  - Blood
UR  - http://dx.doi.org/10.1182/blood-2014-08-594556
VL  - 125
ER  -
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