title: Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.
abstract: A powerful and potentially general approach to the targeting and crystallization of proteins on lipid interfaces through coordination of surface histidine residues to lipid-chelated divalent metal ions is presented. This approach, which should be applicable to the crystallization of a wide range of naturally occurring or engineered proteins, is illustrated here by the crystallization of streptavidin on a monolayer of an iminodiacetate-Cu(II) lipid spread at the air-water interface. This method allows control of the protein orientation at interfaces, which is significant for the facile production of highly ordered protein arrays and for electron density mapping in structural analysis of two-dimensional crystals. Binding of native streptavidin to the iminodiacetate-Cu lipids occurs via His-87, located on the protein surface near the biotin binding pocket. The two-dimensional streptavidin crystals show a previously undescribed microscopic shape that differs from that of crystals formed beneath biotinylated lipids.
authors.affiliation: Center for Bioengineering, University of Washington, Seattle, 98195, USA.
authors.forename: W
authors.initials: W
authors.lastname: Frey
authors.forename: W R
authors.initials: WR
authors.lastname: Schief
authors.forename: D W
authors.initials: DW
authors.lastname: Pack
authors.forename: C T
authors.initials: CT
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Chilkoti
authors.forename: P
authors.initials: P
authors.lastname: Stayton
authors.forename: V
authors.initials: V
authors.lastname: Vogel
authors.forename: F H
authors.initials: FH
authors.lastname: Arnold
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001665:Binding Sites; D002614:Chelating Agents; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine / Q000737:chemistry; D007477:Ions; D008055:Lipids / Q000737:chemistry; D008670:Metals; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D019809:Streptavidin; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D002614:Chelating Agents; D007477:Ions; D008055:Lipids; D008670:Metals; D011506:Proteins; D006639:Histidine; D019809:Streptavidin