title: Generation of functional antibodies for mammalian membrane protein crystallography.
abstract: Membrane proteins act as gateways to cells, and they are responsible for much of the communication between cells and their environments. Crystallography of membrane proteins is often limited by the difficulty of crystallization in detergent micelles. Co-crystallization with antibody fragments has been reported as a method to facilitate the crystallization of membrane proteins; however, it is widely known that the generation of mouse monoclonal antibodies that recognize the conformational epitopes of mammalian integral membrane proteins is typically difficult. Here, we present our protocols to generate functional mouse antibodies for the membrane protein crystallography, which have enabled us to solve crystal structures of mammalian receptors and transporters complexed with antibody fragments.
authors.affiliation: Iwata Human Receptor Crystallography Project, ERATO, JST, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Hino
authors.forename: So
authors.initials: S
authors.lastname: Iwata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Murata
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018360:Crystallography, X-Ray / Q000379:methods*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000276:immunology / Q000378:metabolism; D008081:Liposomes / Q000276:immunology; D026901:Membrane Transport Proteins / Q000276:immunology / Q000648:ultrastructure*; D051379:Mice; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000276:immunology / Q000648:ultrastructure*
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D008081:Liposomes; D026901:Membrane Transport Proteins; D011956:Receptors, Cell Surface