Crystallization and diffraction properties of the Fab fragment of 3B5H10, an antibody specific for disease-causing polyglutamine stretches

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Dec 1;61(Pt 12):1065-8. doi: 10.1107/S1744309105036547. Epub 2005 Nov 24.

Abstract

Because it binds soluble forms of proteins with disease-associated polyglutamine expansions, the antibody 3B5H10 is a powerful tool for studying polyglutamine-related diseases. Crystals of the 3B5H10 Fab (47 kDa) were obtained by vapor diffusion at room temperature from PEG 3350. However, the initial crystals gave highly anisotropic diffraction patterns. After optimization of the crystallization conditions and cryoprotectants, a nearly isotropic diffraction pattern at 2.6 A resolution was achieved for crystals with unit-cell parameters a = 133.26, b = 79.52, c = 41.49 A and space group P2(1)2(1)2. Dehydrated crystals diffracted isotropically to 1.9 A with unit-cell parameters a = 123.65, b = 78.25, c = 42.26 A, beta = 90.3 degrees and space group P2(1).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anisotropy
  • Antibodies / chemistry
  • Crystallization
  • Crystallography, X-Ray
  • Humans
  • Huntingtin Protein
  • Immunoglobulin Fragments / chemistry*
  • Mutation
  • Nerve Tissue Proteins / chemistry
  • Nuclear Proteins / chemistry
  • Peptides / chemistry*
  • Polyethylene Glycols / chemistry
  • Temperature
  • X-Ray Diffraction

Substances

  • Antibodies
  • HTT protein, human
  • Huntingtin Protein
  • Immunoglobulin Fragments
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Peptides
  • polyglutamine
  • Polyethylene Glycols