TY - JOUR T1 - Glutamine 666 renders murine ADAM10 an inefficient <em>S. aureus</em> α-toxin receptor JF - bioRxiv DO - 10.1101/2022.05.11.491455 SP - 2022.05.11.491455 AU - Gisela von Hoven AU - Martina Meyenburg AU - Claudia Neukirch AU - Daniel Siedenschnur AU - Matthias Husmann Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/05/11/2022.05.11.491455.abstract N2 - S. aureus is one of the most important causes of infectious diseases in hospitalized individuals and outpatients. The majority of clinical isolates secretes large amounts of the small membrane pore-forming α-toxin, alias α-hemolysin, which serves as an important virulence factor of this organism. The identification of A Disintegrin And Metalloprotease (ADAM10) as its high affinity receptor held great promise for a better understanding of the processes underlying membrane damage by α-toxin. Twelve years on however, the molecular details of initial toxin binding to target cells remain elusive. Because we noted that several murine cell lines were resilient to α-toxin, we considered the possibility that murine ADAM10 could be less efficient a receptor, as compared to human or bovine orthologues. Accordingly, we sought to identify amino acid residues in ADAM10, which could explain species-dependent functionality as receptor for α-toxin. Our work led to the finding that replacement of a single glutamine residue (Q666) in murine ADAM10 with corresponding glutamic acid (E665) of human or bovine ADAM10 enhances significantly the binding and consequent cytotoxicity of α-toxin. Consistently, a synthetic peptide comprising E665 mitigated α-toxin-dependent hemolysis. In multicellular organisms, E665 is highly conserved, but mice and several other members of the taxon glires evolved glutamine at the corresponding position. The residue is located in a short membrane proximal, extracellular region of ADAM10. Taken together, available structural information, in silico docking, and functional data suggests that α-toxin monomers could bind to cellular membranes via this so-called stalk region of ADAM10 and phosphocholine.Competing Interest StatementThe authors have declared no competing interest.AbbreviationsADAM10,A Disintegrin And Metalloprotease;bADAM10,bovine ADAM10;FACS,fluorescence activated cell sorting;FITC,fluorescein-isothiocyanate;HA,Hemagglutinin;hADAM10,human ADAM10;HAP1ADAM10KO cells,haploid human cell line with knock out of ADAM10 by CRISPR/CAS9;I-TASSER,Iterative Threading ASSEmbly Refinement;LOAD,Late Onset Alzheimer’s Disease;mADAM10,murine ADAM10;PD,prodomain of ADAM10;ΔPD,ADAM10 devoid of the PD;PIpropidium iodide;RRBC,rabbit red blood cells ER -