title: Structural studies examining the substrate specificity profiles of PC-PLC(Bc) protein variants.
abstract: The phosphatidylcholine preferring phospholipase C from Bacillus cereus (PC-PLC(Bc)) catalyzes the hydrolysis of phospholipids in the following order of preference: phosphatidylcholine (PC)>phosphatidylethanolamine (PE)>phosphatidylserine (PS). In previous work, mutagenic, kinetic, and crystallographic experiments suggested that varying the amino acids at the 4th, 56th, and 66th positions had a significant influence upon the substrate specificity profile of PC-PLC(Bc). Here, we report the crystal structures of the native form of several PC-PLC(Bc) variants that exhibited altered substrate specificities for PC, PE, and PS at maximum resolutions of 1.90-2.05 Angstrom. Comparing the structures of these variants to the structure of the wild-type enzyme reveals only minor differences with respect to the number and location of active site water molecules and the side chain conformations of residues at the 4th and 56th positions. These results suggest that subtle changes in steric and electronic properties in the substrate binding site of PC-PLC(Bc) are responsible for the significant changes in substrate selectivity.
authors.affiliation: Department of Chemistry and Biochemistry, The Institute of Cellular and Molecular Biology, The University of Texas, 1 University Station - A5300, Austin, TX 78712, USA.
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Benfield
authors.forename: Nina M
authors.initials: NM
authors.lastname: Goodey
authors.forename: Lauren T
authors.initials: LT
authors.lastname: Phillips
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Martin
mesh_terms: D001409:Bacillus cereus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000737:chemistry* / Q000235:genetics
chemical_list: D020033:Protein Isoforms; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C