Purification, crystallographic analysis of rhesus MHC-I Mamu-A*02 complexed with simian immunodeficiency virus nonapeptide / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1028-1034, 2009.
Article
em Zh
| WPRIM
| ID: wpr-286606
Biblioteca responsável:
WPRO
ABSTRACT
Rhesus macaque (Macaca mulatta) is the best model to study of human immunodeficiency virus (HIV) infection and to develop acquired immunodeficiency syndrome (AIDS) vaccine. The crystal structure of its major histocompatibility antigen complex (MHC) is helpful to understand the mechanism of HIV immune evasion. In this study, we cloned the light chain (beta2m) of MHC class I allele of rhesus macaques, Mamu-A*02, and inserted it into pET21a(+) vector. We transfected the recombinant plasmid pET21a(+)-Mamu-beta2m and pET21a(+)-Mamu-alpha into BL21(DE3). Mamu-A*02 and beta2m were expressed in the form of inclusion bodies in BL21 (DE3). We co-refolded the inclusion bodies of Mamu-alpha and Mamu-beta2m with SIV nonapeptide YY9 and obtained the correct refolded protein complex. Then we purified the protein complex by the gel filtration and anion-exchange column. With hanging-drop method, we screened and optimized for the protein crystal. We managed to collect a X-ray diffraction with the resolution to 2.8 angstroms in the condition of 0.1 mol/L BIS-TRIS (pH5.5), 2.0 mol/L(NH4)2SO4. This crystal belong to perpendicular space group P2(1)2(1)2(1), with unit-cell parameters a = 128.99 angstroms, b = 129.01 angstroms, c = 129.03 angstroms. This data is available for the structure determination.
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Base de dados:
WPRIM
Assunto principal:
Oligopeptídeos
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Antígenos de Histocompatibilidade Classe I
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Vírus da Imunodeficiência Símia
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Cristalografia por Raios X
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Alergia e Imunologia
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Escherichia coli
/
Genética
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Macaca mulatta
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Metabolismo
/
Epitopos
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2009
Tipo de documento:
Article