Preparation and crystallization of a human immunodeficiency virus p24-Fab complex.
Preparation and crystallization of a human immunodeficiency virus p24-Fab complex.
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人类免疫缺陷病毒 p24-Fab 复合物的制备和结晶。
DOI：
10.1073/pnas.87.24.9980
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发表时间：
1990
影响因子：
11.1
通讯作者：
McClure,J
中科院分区：
文献类型：
--
作者：
Prongay,AJ;Smith,TJ;Rossmann,MG;Ehrlich,LS;Carter,CA;McClure,J
在大肠杆菌中表达的重组形式的人免疫缺陷病毒衣壳蛋白p24已被纯化至均一并分离成不同的等电形式。单克隆抗体mAb25.4识别p24氨基末端区域的表位，已从腹水中纯化至均一，并用木瓜蛋白酶消化以产生相应的抗原结合片段（Fab）。从消化混合物中纯化Fab25.4并分离成两种不同的等电形式。通过孵育等摩尔量的两种蛋白质，使两种Fab物质各自与重组p24的一种等电形式复合。用12- 24%PEG 3350作沉淀剂，通过气相扩散法得到了两种不同晶体形态的p24-Fab25.4复合物。这些晶型之一具有a = 92.1 A，B = 85.4 A，c = 54.0 A，α = γ = 90.0度和β = 90.4度的晶胞参数，并且属于单斜晶系空间群P2（1），每个不对称单元具有一个络合物分子。这些晶体强烈地衍射X射线到至少2.7-A分辨率。
A recombinant form of human immunodeficiency virus capsid protein, p24, expressed in Escherichia coli has been purified to homogeneity and separated into distinct isoelectric forms. A monoclonal antibody, mAb25.4, which recognizes an epitope in the amino-terminal region of p24, has been purified to homogeneity from ascites fluid and digested with papain to produce the respective antigen-binding fragment (Fab). The Fab25.4 was purified from the digestion mixture and separated into two distinct isoelectric forms. The two Fab species were each complexed with one isoelectric form of the recombinant p24 by incubating equimolar quantities of the two proteins. Two different crystal morphologies of the p24-Fab25.4 complex were obtained by the vapor-diffusion method with 12-24% PEG 3350 as the precipitant. One of these crystal forms has unit-cell parameters of a = 92.1 A, b = 85.4 A, c = 54.0 A, alpha = gamma = 90.0 degrees and beta = 90.4 degrees and belongs to the monoclinic space group P2(1), with one molecule of the complex per asymmetric unit. These crystals strongly diffracted x-rays to at least 2.7-A resolution.