GPIbα-vWF rolling under shear stress shows differences between type 2B and 2M von Willebrand disease.
Both type 2B and type 2M von Willebrand disease result in bleeding disorders; however, whereas type 2B has increased binding affinity between platelet glycoprotein Ibα and von Willebrand factor (vWF), type 2M has decreased binding affinity between these two molecules. We used R687E type 2B and G561S type 2M vWF-A1 mutations to study binding between flowing platelets and insolubilized vWF mutants. We measured rolling velocities, mean stop times, and mean go times at 37°C using high-speed video microscopy. The rolling velocities for wt-wt interactions first decrease, reach a minimum, and then increase with increasing shear stress, indicating a catch-slip transition. By changing the viscosity, we were able to quantify the effects of force versus shear rate for rolling velocities and mean stop times. Platelet interactions with loss-of-function vWF-A1 retain the catch-slip bond transition seen in wt-wt interactions, but at a higher shear stress compared with the wt-wt transition. The mean stop time for all vWF-A1 molecules reveals catch-slip transitions at different shear stresses (gain-of-function vWF-A1 < wt vWF-A1< loss-of-function vWF-A1). The shift in the catch-slip transition may indicate changes in how the different mutants become conformationally active, indicating different mechanisms leading to similar bleeding characteristics.
Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Emory University School of Medicine, Atlanta, USA.
L A
LA
Coburn
V S
VS
Damaraju
S
S
Dozic
S G
SG
Eskin
M A
MA
Cruz
L V
LV
McIntire
D001792:Blood Platelets / Q000378:metabolism*; D001809:Blood Viscosity; D006801:Humans; D066298:In Vitro Techniques; D008968:Molecular Conformation; D009154:Mutation / Q000235:genetics*; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D013314:Stress, Mechanical; D014842:von Willebrand Diseases / Q000235:genetics*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D019038:Platelet Glycoprotein GPIb-IX Complex; D014841:von Willebrand Factor
J Thromb Haemost. 2006 Oct;4(10):2103-14
16889557
Biophys J. 2007 Feb 1;92(3):787-97
17085490
Nature. 2003 May 8;423(6936):190-3
12736689
J Biol Chem. 1993 Oct 5;268(28):21238-45
8407961
Science. 2002 Aug 16;297(5584):1176-9
12183630
Cell. 1998 Sep 4;94(5):657-66
9741630
J Biol Chem. 1995 May 5;270(18):10822-7
7738019
J Thromb Haemost. 2006 Feb;4(2):417-25
16420575
Biochem J. 1997 May 15;324 ( Pt 1):185-91
9164855
J Biol Chem. 1998 Apr 24;273(17):10396-401
9553097
J Biol Chem. 1995 Jun 2;270(22):13406-14
7539426
Blood. 1987 Nov;70(5):1577-83
2889486
Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9846-9
1409710
Biophys J. 2009 Jul 22;97(2):618-27
19619477
J Thromb Haemost. 2006 Oct;4(10):2151-61
16805860
J Clin Invest. 2008 Sep;118(9):3195-207
18725999
J Mol Biol. 2007 Feb 23;366(3):986-1000
17187823
J Biol Chem. 2004 Jan 16;279(3):2291-8
14573602
Annu Rev Med. 2002;53:75-88
11818464
Thromb Haemost. 1996 Jun;75(6):950-8
8822592
Biophys J. 2003 Dec;85(6):4099-109
14645097
Biophys J. 2002 Jul;83(1):194-205
12080112
Proc R Soc Lond B Biol Sci. 1988 Jun 22;234(1274):55-83
2901109
J Cell Biol. 1997 Sep 8;138(5):1169-80
9281593
Int J Biochem Cell Biol. 1997 Jan;29(1):91-105
9076944
Blood. 2002 Dec 1;100(12):4033-9
12393397
J Cell Biol. 2004 Sep 13;166(6):913-23
15364963
J Biol Chem. 2000 Jun 23;275(25):19098-105
10764791
Haemophilia. 2008 Nov;14 Suppl 5:11-26
18786007
Thromb Haemost. 1998 Jan;79(1):211-6
9459349