Crystal structure of phospholipase C from Bacillus cereus complexed with a substrate analog.
We report the first crystal structure of a complex between PLC from Bacillus cereus (PLCBc) and a competitive inhibitor that is an analog of the natural phospholipid substrate. The structure has been determined at 1.9 A resolution and refined to a final R-factor of 15.7%. The inhibitor binds with its phosphonyl group to the three Zn ions in the active site of the enzyme and is also involved in a hydrogen bonded network including several water molecules and amino acid side-chains which appear to help orient the substrate for productive binding. The interactions within this complex provide some important information regarding the mechanism of PLC-catalyzed hydrolysis of membrane phospholipids. A water molecule, located approximately apical to the diacylglycerol leaving group, seems to be the most likely candidate for the attacking nucleophile which initiates the reaction.
Hansen
S
S
Institute of Mathematical and Physical Sciences, University of Tromsø, Norway.
Hough
E
E
Svensson
L A
LA
Wong
Y L
YL
Martin
S F
SF
D001409:Bacillus cereus / Q000201:enzymology*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D008024:Ligands; D008958:Models, Molecular; D010713:Phosphatidylcholines / Q000737:chemistry; D017434:Protein Structure, Tertiary; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000648:ultrastructure*
D008024:Ligands; D010713:Phosphatidylcholines; D010738:Type C Phospholipases