Crystallization and X-ray analysis of the Y75N mutant of Mucor pusillus pepsin complexed with inhibitor.
Y75N mutant Mucor pusillus pepsin has been overexpressed in yeast, purified and cocrystallized with the iodine-containing human renin inhibitor CP-113972 [(2R,3S]-isopropyl 3-[(L-prolyl-p-iodo-L-phenylalanyl-S-methyl-cysteinyl)amino-4]-cyclohexyl-2-hydroxybutanoate] for X-ray crystallography. Tetragonal complex crystals with space group P4(3)2(1)2 were produced by the hanging-drop vapour-diffusion method and diffracted to 3.0 A. The crystals exhibited unit-cell parameters a = b = 182.5, c = 99.1 A and contained four molecules in the asymmetric unit. A 96% complete data set was collected at 298 K using Cu Kalpha X-rays from a rotating-anode generator. Solution of the crystal structure of Y75N mutant M. pusillus pepsin is under way by molecular replacement using the molecular coordinates of wild-type M. pusillus pepsin as a model.
Badasso
Mohammed O
MO
Department of Oral Science, University of Minnesota, Minneapolis, MN 55455, USA. badas001@umn.edu
Dhanaraj
V
V
Wood
S P
SP
Cooper
J B
JB
Blundell
T L
TL
D002087:Butyrates / Q000737:chemistry; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry; D009089:Mucor; D020125:Mutation, Missense; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding
D002087:Butyrates; C117374:CP 113972; D010434:Pepsin A; D003545:Cysteine