Crystallization of Pichia pastoris lysyl oxidase.
A copper-containing amine oxidase (PPLO) from the yeast Pichia pastoris has been purified and crystallized in two forms. PPLO is a glycoprotein. The molecular mass from SDS-polyacrylamide gels is 112 kDa, consistent with 20% glycosylation by weight (the calculated molecular weight of the polypeptide is 89.7 kDa). Orthorhombic crystals belonging to space group P2(1)2(1)2(1), with unit-cell parameters a = 163.7, b = 316.1, c = 84.0 A, diffract to 2.65 A resolution. Monoclinic crystals belonging to space group C2, with unit-cell parameters a = 248.4, b = 121.1, c = 151.8 A, beta = 124.6 degrees, diffract to 1.65 A resolution. Native data have been recorded from each crystal form at 100 K using synchrotron radiation. A self-rotation function for the monoclinic crystal form reveals the presence of a non-crystallographic twofold axis perpendicular to the crystallographic twofold axis, consistent with the presence of two dimers in the asymmetric unit.
Lee
Mihwa
M
School of Molecular and Microbial Biosciences, University of Sydney, NSW 2006, Australia.
Willingham
Katrina
K
Langley
David
D
Maher
Megan J
MJ
Cohen
Aina E
AE
Ellis
Paul J
PJ
Kuchar
Jason A
JA
Dooley
David M
DM
Freeman
Hans C
HC
Guss
J Mitchell
JM
D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D010843:Pichia / Q000201:enzymology*; D011487:Protein Conformation; D008249:Protein-Lysine 6-Oxidase / Q000737:chemistry* / Q000302:isolation & purification
D008249:Protein-Lysine 6-Oxidase