The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors
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...
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MDPI AG
2022-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/5/2440 |
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| author | Chek Ziu Koo Alexandra L. Matthews Neale Harrison Justyna Szyroka Bernhard Nieswandt Elizabeth E. Gardiner Natalie S. Poulter Michael G. Tomlinson |
| author_facet | Chek Ziu Koo Alexandra L. Matthews Neale Harrison Justyna Szyroka Bernhard Nieswandt Elizabeth E. Gardiner Natalie S. Poulter Michael G. Tomlinson |
| author_sort | Chek Ziu Koo |
| collection | DOAJ |
| description | 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. |
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| id | doaj.art-87659b7c4198463b847b3ff79a0e3b38 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T20:39:03Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-87659b7c4198463b847b3ff79a0e3b382023-11-23T23:04:02ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-02-01235244010.3390/ijms23052440The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular ScissorsChek Ziu Koo0Alexandra L. Matthews1Neale Harrison2Justyna Szyroka3Bernhard Nieswandt4Elizabeth E. Gardiner5Natalie S. Poulter6Michael G. Tomlinson7School of Biosciences, University of Birmingham, Birmingham B15 2TT, UKSchool of Biosciences, University of Birmingham, Birmingham B15 2TT, UKSchool of Biosciences, University of Birmingham, Birmingham B15 2TT, UKSchool of Biosciences, University of Birmingham, Birmingham B15 2TT, UKInstitute of Experimental Biomedicine I, University Hospital and Rudolf Virchow Center Würzburg, University of Würzburg, D-97080 Würzburg, GermanyDivision of Genome Science and Cancer, John Curtin School of Medical Research, Australian National University, Canberra ACT 2601, AustraliaCentre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Midlands B15 2TT, UKSchool of Biosciences, University of Birmingham, Birmingham B15 2TT, UKThe 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.https://www.mdpi.com/1422-0067/23/5/2440ADAM10GPVItetraspaninplateletsheddingTspanC8 |
| spellingShingle | Chek Ziu Koo Alexandra L. Matthews Neale Harrison Justyna Szyroka Bernhard Nieswandt Elizabeth E. Gardiner Natalie S. Poulter Michael G. Tomlinson The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors International Journal of Molecular Sciences ADAM10 GPVI tetraspanin platelet shedding TspanC8 |
| title | The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors |
| title_full | The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors |
| title_fullStr | The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors |
| title_full_unstemmed | The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors |
| title_short | The Platelet Collagen Receptor GPVI Is Cleaved by Tspan15/ADAM10 and Tspan33/ADAM10 Molecular Scissors |
| title_sort | platelet collagen receptor gpvi is cleaved by tspan15 adam10 and tspan33 adam10 molecular scissors |
| topic | ADAM10 GPVI tetraspanin platelet shedding TspanC8 |
| url | https://www.mdpi.com/1422-0067/23/5/2440 |
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