title: Electron microscopy of biotinylated protein complexes bound to streptavidin monolayer crystals.
abstract: Chemical biotinylation of protein complexes followed by binding to two-dimensional (monolayer) crystals of streptavidin is shown to be an effective way to prepare cryo-EM specimens from samples at low protein concentration. Three different multiprotein complexes are used to demonstrate the generality of this method. In addition, native thermosomes, purified from Sulfolobus solfataricus P2, are used to demonstrate that a uniform distribution of Euler angles is produced, even though this particle is known to adopt a preferred orientation when other methods of cryo-EM specimen preparation are used.
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA.
authors.forename: Bong-Gyoon
authors.initials: BG
authors.lastname: Han
authors.forename: Ross W
authors.initials: RW
authors.lastname: Walton
authors.forename: Amos
authors.initials: A
authors.lastname: Song
authors.forename: Peter
authors.initials: P
authors.lastname: Hwu
authors.forename: Milton T
authors.initials: MT
authors.lastname: Stubbs
authors.forename: Steven M
authors.initials: SM
authors.lastname: Yannone
authors.forename: Pablo
authors.initials: P
authors.lastname: Arbeláez
authors.forename: Ming
authors.initials: M
authors.lastname: Dong
authors.forename: Robert M
authors.initials: RM
authors.lastname: Glaeser
mesh_terms: D000327:Adsorption; D000818:Animals; D001052:Apoferritins / Q000737:chemistry / Q000648:ultrastructure; D001426:Bacterial Proteins / Q000737:chemistry; D001710:Biotin / Q000737:chemistry*; D019921:Biotinylation; D020285:Cryoelectron Microscopy / Q000379:methods*; D003460:Crystallization; D016969:Desulfovibrio vulgaris; D006736:Horses; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000737:chemistry / Q000648:ultrastructure; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D019809:Streptavidin / Q000737:chemistry*; D048229:Sulfolobus solfataricus; D056405:Thermosomes / Q000737:chemistry / Q000648:ultrastructure
chemical_list: D001426:Bacterial Proteins; D009097:Multienzyme Complexes; D001710:Biotin; C044822:6,7-dimethyl-8-ribityllumazine synthase; D019809:Streptavidin; D001052:Apoferritins; D056405:Thermosomes