Five viral peptide-HLA-A2 co-crystals. Simultaneous space group determination and X-ray data collection.
We prepared and crystallized five complexes of the human histocompatibility molecule HLA-A2 with peptides derived from human immunodeficiency virus type 1, human T lymphotropic virus type 1, influenza A virus and hepatitis B virus proteins. Each HLA-A2 complex was refolded in vitro from insoluble proteins produced in bacteria; to crystallize, two of the complexes required seeding with microcrystals of another complex. Maintained at -160 degrees C, single co-crystals of each of the five peptide-HLA-A2 complexes yielded complete X-ray diffraction data sets to a resolution of approximately 2.5 A. After a sufficient number of diffraction peaks were acquired during data collection, the direct analysis of integrated intensities established the point group of the co-crystal, thus allowing an efficient data collection strategy to be designed. The subsequent examination of systematic absences revealed that the five peptide-HLA-A2 co-crystals formed in space groups P1, P2(1), or P2(1)2(1)2(1). Molecular replacement structure solutions yielded unambiguous protein electron density maps, thus confirming the space group determinations. The system of obtaining HLA-A2 co-crystal structures described here is applicable to other crystallographic problems where structures of several related molecules from uncharacterized single crystals are required.
Garboczi
D N
DN
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Madden
D R
DR
Wiley
D C
DC
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000737:chemistry; D015789:HLA-A2 Antigen / Q000737:chemistry* / Q000378:metabolism; D006515:Hepatitis B virus / Q000737:chemistry; D006801:Humans; D009980:Influenza A virus / Q000737:chemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry; D014758:Viral Core Proteins / Q000737:chemistry; D014763:Viral Matrix Proteins / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D004268:DNA-Binding Proteins; D015789:HLA-A2 Antigen; D009842:Oligopeptides; D014758:Viral Core Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C080957:tax-responsive element-binding protein 302; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase