title: In vitro screening for molecules that affect virus capsid assembly (and other protein association reactions).
abstract: Protein self-assembly is critical for numerous biological processes. Yet, assembly is rarely targeted by therapeutic agents, in part because it is hard to identify molecules that interfere with protein-protein interactions. Here, we describe a simple fluorescence-based screen for self-association and its application to the assembly of hepatitis B virus capsids. These data are analyzed to identify kinetic and thermodynamic effects--both of which are critical for the viral lifecycle and for understanding the mechanism of assembly effectors. Suggestions are made for modification of this protocol so that it can be applied to other self-assembling systems. With manual pipetting, setting up a plate takes about 2 h, the initial reading takes 1 h and the end point reading the following day takes about 5 min.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA. adam-zlotnick@ouhsc.edu
authors.forename: Adam
authors.initials: A
authors.lastname: Zlotnick
authors.forename: Angela
authors.initials: A
authors.lastname: Lee
authors.forename: Christina R
authors.initials: CR
authors.lastname: Bourne
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Johnson
authors.forename: Paul L
authors.initials: PL
authors.lastname: Domanico
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Stray
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002213:Capsid / Q000187:drug effects* / Q000502:physiology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005453:Fluorescence; D006515:Hepatitis B virus / Q000187:drug effects* / Q000502:physiology; D066298:In Vitro Techniques; D007700:Kinetics; D008954:Models, Biological; D019065:Virus Assembly / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents