Purification, crystallization and preliminary X-ray crystallographic analysis of recombinant murine Golgi mannosidase IA, a class I alpha-mannosidase involved in Asn-linked oligosaccharide maturation

Acta Crystallogr D Biol Crystallogr. 1999 Feb;55(Pt 2):571-3. doi: 10.1107/s090744499801244x.

Abstract

Golgi mannosidase IA is a class I alpha-mannosidase which catalyzes the conversion of Man9GlcNAc2 or Man8GlcNAc2 oligosaccharide substrates to Man5GlcNAc2 during the maturation of Asn-linked oligosaccharides. The enzyme is a type II membrane protein, and a recombinant form of mannosidase IA from mouse, lacking the transmembrane domain, has been expressed in Pichia pastoris, purified to homogeneity and crystallized by the hanging-drop vapor-diffusion method. The crystals grow as thin rods, with unit-cell dimensions a = 54.9, b = 135.01, c = 69.9 A. The crystals exhibit the symmetry of space group P2221 and diffract to 2.8 A resolution. The asymmetric unit contains one monomer ( approximately 53 kDa) and has a solvent content of 59%.

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

  • Animals
  • Asparagine / chemistry*
  • Crystallization
  • Crystallography, X-Ray
  • Golgi Apparatus / enzymology*
  • Isoenzymes / chemistry*
  • Mannosidases / chemistry*
  • Mice
  • Oligosaccharides / chemistry*
  • Protein Conformation
  • Recombinant Proteins / chemistry

Substances

  • Isoenzymes
  • Oligosaccharides
  • Recombinant Proteins
  • Asparagine
  • Mannosidases
  • mannosyl-oligosaccharide 1,2-alpha-mannosidase