Projects per year
Abstract
The platelet-activating collagen receptor GPVI represents the focus of clinical trials as an antiplatelet target for arterial thrombosis, and soluble GPVI is a plasma biomarker for several human diseases. A disintegrin and metalloproteinase 10 (ADAM10) acts as a 'molecular scissor' that cleaves the extracellular region from GPVI and many other substrates. ADAM10 interacts with six regulatory tetraspanin membrane proteins, Tspan5, Tspan10, Tspan14, Tspan15, Tspan17 and Tspan33, which are collectively termed the TspanC8s. These are emerging as regulators of ADAM10 substrate specificity. Human platelets express Tspan14, Tspan15 and Tspan33, but which of these regulates GPVI cleavage remains unknown. To address this, CRISPR/Cas9 knockout human cell lines were generated to show that Tspan15 and Tspan33 enact compensatory roles in GPVI cleavage, with Tspan15 bearing the more important role. To investigate this mechanism, a series of Tspan15 and GPVI mutant expression constructs were designed. The Tspan15 extracellular region was found to be critical in promoting GPVI cleavage, and appeared to achieve this by enabling ADAM10 to access the cleavage site at a particular distance above the membrane. These findings bear implications for the regulation of cleavage of other ADAM10 substrates, and provide new insights into post-translational regulation of the clinically relevant GPVI protein.
Original language | English |
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Article number | 2440 |
Journal | International Journal of Molecular Sciences |
Volume | 23 |
Issue number | 5 |
DOIs | |
Publication status | Published - 23 Feb 2022 |
Keywords
- ADAM10 Protein/metabolism
- Amyloid Precursor Protein Secretases/metabolism
- Blood Platelets/metabolism
- CRISPR-Cas Systems
- Gene Knockout Techniques
- HEK293 Cells
- Humans
- Membrane Proteins/metabolism
- Platelet Membrane Glycoproteins/genetics
- Protein Domains
- Proteolysis
- Substrate Specificity
- Tetraspanins/chemistry
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Dive into the research topics of 'The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors'. Together they form a unique fingerprint.Projects
- 2 Finished
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Identifying the tetraspanin/ADAM10 ‘molecular scissor’ for the platelet collagen and fibrin receptor GPVI
Poulter, N. (Co-Investigator) & Tomlinson, M. (Principal Investigator)
7/05/18 → 6/09/21
Project: Research
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Understanding the mechanism by which tetraspanins regulate the 'molecular scissor' ADAM10
Dafforn, T. (Co-Investigator), Tomlinson, M. (Principal Investigator) & Gough, R. (Co-Investigator)
Biotechnology & Biological Sciences Research Council
1/07/17 → 30/06/20
Project: Research Councils