Crystallization and preliminary X-ray diffraction study of a bacterially produced T-cell antigen receptor V alpha domain.
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.
Fields
B A
BA
Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute.
Ysern
X
X
Poljak
R J
RJ
Shao
X
X
Ward
E S
ES
Mariuzza
R A
RA
D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D046911:Macromolecular Substances; D051379:Mice; D004676:Myelin Basic Protein / Q000378:metabolism; D017510:Protein Folding; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; D004676:Myelin Basic Protein; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins