RT Journal Article SR Electronic T1 Hot-spots and their contribution to the self-assembly of the viral capsid: in-vitro validation JF bioRxiv FD Cold Spring Harbor Laboratory SP 724146 DO 10.1101/724146 A1 Valdez-Lara, Alejandra Gabriela A1 Andrade-Medina, Mariana A1 Alemán-Vilis, Josué Alejandro A1 Pérez-Montoya, Aldo Adrián A1 Pineda-Aguilar, Nayely A1 Martínez-Guerra, Eduardo A1 Gaytán, Paul A1 Carrillo-Tripp, Mauricio YR 2019 UL http://biorxiv.org/content/early/2019/08/06/724146.abstract AB The viral capsid is a macromolecular complex formed by self-assembled proteins which, in many cases, are biopolymers with an identical amino acid sequence. Specific protein-protein interactions drive the capsid self-assembly process. However, it is believed that only a small set of interface residues significantly contribute to the formation of the capsid, the so-called “hot-spots”. Here, we investigate the effect of point-mutations on previously predicted hot-spots of the icosahedral Cowpea Chlorotic Mottle Virus. We characterize the formation and stability of virus-like particles in-vitro by thermal shift assays. Our results show that a single mutation on residue E176 or V189 significantly perturbs the quaternary protein-protein interaction, preventing the formation of the capsid. Furthermore, these mutations do not destroy the tertiary fold of the capsid protein. Our findings give evidence of the in-silico hot-spot prediction accuracy. As a whole, our methodology provides a guide to the rational development of molecules that could inhibit virus formation.CCMVCowpea Chlorotic Mottle VirusCPcapsid proteinVLPvirus-like particlePPIprotein-protein interactionsWTwild-typeCOMcenter-of-mass