Purification, crystallization and preliminary X-ray analysis of the galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) of the ABC transporter from Bifidobacterium longum JCM1217

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Sep 1;63(Pt 9):751-3. doi: 10.1107/S1744309107036263. Epub 2007 Aug 10.

Abstract

A recombinant galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) from Bifidobacterium longum JCM1217 has been prepared and crystallized by the hanging-drop vapour-diffusion method using 10 mg ml(-1) purified enzyme, 0.01 M zinc sulfate, 0.1 M MES buffer pH 5.9-6.4 and 20-22%(v/v) PEG MME 550 in the presence of 5 mM disaccharide ligands. Suitable crystals grew after 10 d incubation at 293 K. The crystals belong to space group C222(1), with unit-cell parameters a = 106.3, b = 143.6, c = 114.6 A for the lacto-N-biose I complex and a = 106.4, b = 143.4, c = 115.5 A for the galacto-N-biose complex, and diffracted to 1.85 and 1.99 A resolution, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry*
  • ATP-Binding Cassette Transporters / isolation & purification
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification
  • Bifidobacterium / chemistry*
  • Crystallization
  • DNA Primers
  • Plasmids
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • X-Ray Diffraction

Substances

  • ATP-Binding Cassette Transporters
  • Bacterial Proteins
  • DNA Primers
  • Recombinant Proteins