Crystallization of Pichia pastoris lysyl oxidase

Acta Crystallogr D Biol Crystallogr. 2002 Dec;58(Pt 12):2177-9. doi: 10.1107/s0907444902016827. Epub 2002 Nov 23.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Crystallization
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Weight
  • Pichia / enzymology*
  • Protein Conformation
  • Protein-Lysine 6-Oxidase / chemistry*
  • Protein-Lysine 6-Oxidase / isolation & purification

Substances

  • Protein-Lysine 6-Oxidase