Structural, Functional and Mechanistic Insights Uncover the Fundamental Role of Orphan Connexin-62 in Platelets

Sahli, Khaled, Flora, Gagan, Sasikumar, Parvathy, Maghrabi, Ali, Holbrook, Lisa, AlOuda, Sarah, Elgheznawy, Amro, Sage, Tanya, Stainer, Alexander, Adiyaman, Recep, AboHassan, Mohammad, Crescente, Marilena, Kriek, Neline, Vayiyapuri, Sakhtivel, Bye, Alexander, Unsworth, Amanda ORCID: https://orcid.org/0000-0003-3809-5984, Jones, Christopher, McGuffin, Liam and Gibbins, Jonathan (2021) Structural, Functional and Mechanistic Insights Uncover the Fundamental Role of Orphan Connexin-62 in Platelets. Blood, 137 (6). pp. 830-843. ISSN 0006-4971

Preview

Accepted Version Available under License In Copyright. Download (10MB) | Preview

Abstract

Connexins (Cxs) oligomerise to form hexameric hemichannels in the plasma membrane that can further dock together on adjacent cells to form gap junctions and facilitate intercellular-trafficking of molecules. In this study, we report the expression and function of an ‘orphan’ connexin, Cx62, in human and mouse (Cx57, mouse homologue) platelets. A novel mimetic peptide (62Gap27) was developed to target the second extracellular loop of Cx62 and 3D structural models predicted its interference with gap junction and hemichannel function. The ability of 62Gap27 to regulate both gap junction and hemichannel-mediated intercellular communication was observed using FRAP analysis and flow cytometry. Cx62 inhibition by 62Gap27 suppressed a range of agonist-stimulated platelet functions and impaired thrombosis and haemostasis. This was associated with elevated PKA-dependent signalling in a cyclic adenosine monophosphate-independent manner, and was not observed in Cx57 deficient mouse platelets (in which the selectivity of 62Gap27 for this connexin was also confirmed). Notably, Cx62 hemichannels were observed to function independently of Cx37 and Cx40 hemichannels. Together, our data reveal a fundamental role for a hitherto uncharacterised connexin in the regulation of the function of circulating cells.