Crystals of an antibody Fv fragment against an integral membrane protein diffracting to 1.28 A resolution

Proteins. 1995 Jan;21(1):74-7. doi: 10.1002/prot.340210110.

Abstract

The Fv fragment of a monoclonal antibody, 7E2 (IgG1, kappa, murine), which is directed against the integral membrane protein cytochrome c oxidase (EC 1.9.3.1) from Paracoccus denitrificans, was cloned and produced in Escherichia coli. Crystals suitable for high-resolution X-ray analysis were obtained by microdialysis under low salt conditions. The crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of a = 51.51 A, b = 56.15 A, c = 99.86 A (1 A = 0.1 nm) and contain one Fv fragment per asymmetric unit. Using synchrotron radiation diffraction data were collected up to 1.28 A resolution. This high resolution is very unusual for a heterodimeric protein. The crystals should open the way for refining not only the atomic positions, but also for obtaining information about internal dynamics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatography, Affinity
  • Cloning, Molecular
  • Crystallization*
  • DNA, Complementary
  • Electron Transport Complex IV / chemistry
  • Electron Transport Complex IV / immunology*
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Immunoglobulin Fragments / chemistry*
  • Immunoglobulin Variable Region / immunology*
  • Mice
  • Molecular Sequence Data
  • Paracoccus denitrificans / chemistry
  • Paracoccus denitrificans / enzymology
  • Paracoccus denitrificans / immunology
  • Plasmids
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • X-Ray Diffraction*

Substances

  • DNA, Complementary
  • Immunoglobulin Fragments
  • Immunoglobulin Variable Region
  • Recombinant Proteins
  • Electron Transport Complex IV