Crystallization of murine major histocompatibility complex class I H-2Kb with single peptides.
X-ray quality crystals of a soluble murine class I H-2Kb molecule complexed with three different peptide antigens were grown in several forms by streak seeding and macroseeding methods. Co-crystals with VSV-8 (RGYVYGQL), OVA-8 (SIINFEKL) and SEV-9 (FAPGNYPAL) peptides were grown either from NaH2PO4/HPO4 or from polyethylene glycol 4000 within the pH range 5.0 to 7.5, with the use of 4-methyl-2-pentane diol (MPD) as an additive. The VSV-8 crystals grew in space groups P1, with cell dimensions a = 63.1 A, b = 69.1 A, c = 72.0 A, alpha = 89.9 degrees, beta = 77.1 degrees, gamma = 123.3 degrees and P2(1)2(1)2, with a = 138.1 A, b = 88.6 A, c = 45.7 A, and diffract to 2.9 and 2.3 A, respectively. Crystals of the SEV-9 complex grew from similar crystallization conditions to those of the orthorhombic VSV-8 complex with similar cell parameters and diffract to at least 2.5 A resolution. Crystals of the OVA-8 complex were obtained from either phosphate (space group C2, a = 118.7 A, b = 61.6 A, c = 85.3 A, beta = 108.4 degrees) or polyethylene glycol (space group P1, a = 64.5 A, b = 71.0 A, c = 66.3 A, alpha = 89.7 degrees, beta = 95.7 degrees, gamma = 123.3 degrees) and diffract to 3 A resolution. The crystallization procedures used here significantly increased the rate and production of X-ray quality crystals.
Stura
E A
EA
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.
Matsumura
M
M
Fremont
D H
DH
Saito
Y
Y
Peterson
P A
PA
Wilson
I A
IA
D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D005805:Genes, MHC Class I; D006183:H-2 Antigens / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D046911:Macromolecular Substances; D008285:Major Histocompatibility Complex; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D014961:X-Ray Diffraction
D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D008024:Ligands; D046911:Macromolecular Substances; D010455:Peptides