Crystallization and preliminary X-ray diffraction study of a bacterially produced T-cell antigen receptor V alpha domain

J Mol Biol. 1994 Jun 3;239(2):339-41. doi: 10.1006/jmbi.1994.1373.

Abstract

A recombinant form of the variable domain of the alpha chain of a murine T-cell receptor specific for the N-terminal nonapeptide of myelin basin protein in association with the major histocompatibility complex class II I-Au molecule has been crystallized in a form suitable for X-ray diffraction analysis. This protein was secreted into the periplasmic space of Escherichia coli cells and affinity-purified using a nickel chelate adsorbent. The crystals are orthorhombic, space group P2(1)2(1)2, with unit cell dimensions a = 97.7 A, b = 79.6 A, c = 30.4 A and diffract to beyond 2.2 A resolution. The ability to crystallize a T-cell receptor domain produced in bacteria strongly suggests that the periplasmic space can provide a suitable environment for the correct in vivo folding of this class of antigen recognition molecules.

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
  • Binding Sites
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray / methods
  • Escherichia coli
  • Histocompatibility Antigens Class II / metabolism
  • Macromolecular Substances
  • Mice
  • Myelin Basic Protein / metabolism
  • Protein Folding
  • Receptors, Antigen, T-Cell / biosynthesis
  • Receptors, Antigen, T-Cell / chemistry*
  • Receptors, Antigen, T-Cell / isolation & purification
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification

Substances

  • Histocompatibility Antigens Class II
  • Macromolecular Substances
  • Myelin Basic Protein
  • Receptors, Antigen, T-Cell
  • Recombinant Proteins