title: A-Disintegrin-And-Metalloproteinase (ADAM) 10 Activity on Resting and Activated Platelets.
abstract: The primary platelet collagen receptor, glycoprotein VI (GPVI), plays an important role in platelet activation and thrombosis. The ectodomain of human GPVI (sGPVI) is proteolytically shed from human platelets by a-disintegrin-and-metalloproteinase 10 (ADAM10). In this study, we used a novel ADAM10-sensitive fluorescence resonance energy transfer sensor to analyze ADAM10-mediated shedding of GPVI from human platelets in response to the exposure of GPVI ligands collagen-related peptide (10 μg/mL), collagen (10 μg/mL), and convulxin (0.1 μg/mL) to shear stress (1000-10000 s(-1), 5 min), or a generic activator of metalloproteinases, N-ethylmaleimide (NEM, 5 mM). Elevated shear, NEM, or ligand engagement of GPVI all induced shedding of GPVI, as detected by release of sGPVI; however, only shear or NEM significantly increased ADAM10 enzyme activity. ADAM10 activity was also detectable on the surface of thrombi formed on a collagen-coated surface under flow conditions. Our findings indicate different mechanisms regulate shear- and ligand-induced shedding and shear forces found within the vasculature can regulate ADAM10 activity.
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Adam
authors: A
authors: Facey
authors: Department of Mechanical and Aerospace Engineering, Monash University , Clayton, Victoria, Australia 3168.
authors: Isaac
authors: I
authors: Pinar
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Jane F
authors: JF
authors: Arthur
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Jianlin
authors: J
authors: Qiao
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Jing
authors: J
authors: Jing
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Belden
authors: B
authors: Mado
authors: Department of Mechanical and Aerospace Engineering, Monash University , Clayton, Victoria, Australia 3168.
authors: Josie
authors: J
authors: Carberry
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Robert K
authors: RK
authors: Andrews
authors: Australian Centre for Blood Diseases, Monash University , Melbourne, Victoria, Australia 3004.
authors: Elizabeth E
authors: EE
authors: Gardiner
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D001777:Blood Coagulation; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D003435:Crotalid Venoms / Q000378:metabolism; D019483:Disintegrins / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D015539:Platelet Activation; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D013927:Thrombosis / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D003435:Crotalid Venoms; D019483:Disintegrins; D037181:Lectins, C-Type; D008565:Membrane Proteins; D010455:Peptides; D010980:Platelet Membrane Glycoproteins; C435908:collagen-related peptide; C098341:platelet membrane glycoprotein VI; C028890:convulxin; D003094:Collagen; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human