title: Structural characterization of ordered arrays of sn-glycerol-3-phosphate acyltransferase from Escherichia coli.
abstract: Overproduction of the sn-glycerol-3-phosphate acyltransferase in Escherichia coli leads to incorporation of this integral membrane protein into ordered tubular arrays within the cell. Freeze-fracture-etch shadowing was performed on suspensions of partially purified tubules and whole bacteria. This procedure revealed the presence of ridges and grooves defining a set of long-pitch left-handed helical ridges. The long-pitch helices represented chains of acyltransferase dimers. Tubules observed within the cell were often closely packed, with an apparent alignment of grooves and ridges in adjacent tubules. Fracture planes passing through the tubules indicated the presence of a bilayer structure, with some portion of the enzyme being associated with the membrane. The major portion of the enzyme extended from the hydrophilic surface, forming a large globular structure that, in favorable views, displayed a central cavity facing the cytoplasm. Computer analysis of shadowed tubules revealed that the left-handed helices were six stranded, with a pitch of 1,050 A (105.0 nm) and a spacing of 75 A (7.5 nm) between acyltransferase dimers along the chains. Analysis of the predicted secondary structure failed to reveal obvious transmembrane segments, suggesting that very little of the protein was inserted into the bilayer.
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
authors.forename: W O
authors.initials: WO
authors.lastname: Wilkison
authors.forename: R M
authors.initials: RM
authors.lastname: Bell
authors.forename: K A
authors.initials: KA
authors.lastname: Taylor
authors.forename: M J
authors.initials: MJ
authors.lastname: Costello
mesh_terms: D003593:Cytoplasm / Q000201:enzymology / Q000648:ultrastructure; D004926:Escherichia coli / Q000201:enzymology* / Q000648:ultrastructure; D005613:Freeze Etching; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000737:chemistry / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning
chemical_list: D005992:Glycerol-3-Phosphate O-Acyltransferase