Kaufman J, Salomonsen J, Skjødt K, Thorpe D
Basel Institute for Immunology, Switzerland.
Proc Natl Acad Sci U S A. 1990 Nov;87(21):8277-81. doi: 10.1073/pnas.87.21.8277.
B-G antigens are cell-surface molecules encoded by a highly polymorphic multigene family located in the chicken major histocompatibility complex (MHC). Rabbit antisera to B-G molecules immunoprecipitate 3-6 bands from iodinated erythrocytes by sodium dodecyl sulfate (SDS) gels under reducing conditions. These are all B-G molecules because they all map to the B-G region of the chicken MHC in congenic and recombinant chickens, most are directly recognized by the antisera, most form disulfide-linked dimers, and none bear N-linked carbohydrate. Both apparent homodimers and heterodimers are found, which bear intrachain disulfide bonds. All 3-6 bands have different mobilities in SDS gels between different haplotypes, ranging from 30 to 55 kDa. This size polymorphism is not affected by glycosidase treatment or addition of protease inhibitors. Partial proteolysis of cell surface-iodinated B-G molecules generates extremely similar patterns of spots, both within and between haplotypes. These surface-iodinated peptides bear either interchain or intrachain disulfide bonds. Additional peptides are generated by proteolysis of B-G molecules iodinated after isolation. Thus, it appears that the extracellular regions of these molecules are very similar and that the length polymorphism is due to variations in the cytoplasmic regions. Inspection of the cDNA-derived protein sequence in this region shows many heptad repeats, which may allow variation in length by step deletion and alternative splicing. The repeats indicate an alpha-helical coiled-coil structure, which could form an interaction between subunits of the dimer or with the cytoskeleton or both.
B - G抗原是由位于鸡主要组织相容性复合体(MHC)中的一个高度多态的多基因家族编码的细胞表面分子。在还原条件下,用十二烷基硫酸钠(SDS)凝胶对碘化红细胞进行免疫沉淀时,针对B - G分子的兔抗血清可沉淀出3 - 6条带。这些都是B - G分子,因为在同基因和重组鸡中它们都定位到鸡MHC的B - G区域,大多数能被抗血清直接识别,大多数形成二硫键连接的二聚体,且都不带有N - 连接的碳水化合物。发现了明显的同型二聚体和异型二聚体,它们都带有链内二硫键。在不同单倍型之间,所有这3 - 6条带在SDS凝胶中的迁移率都不同,范围从30到55 kDa。这种大小多态性不受糖苷酶处理或添加蛋白酶抑制剂的影响。细胞表面碘化的B - G分子经部分蛋白酶解后,在单倍型内部和之间都产生极其相似的斑点模式。这些表面碘化的肽带有链间或链内二硫键。分离后碘化的B - G分子经蛋白酶解还会产生其他肽段。因此,这些分子的细胞外区域似乎非常相似,长度多态性是由于细胞质区域的变化所致。对该区域cDNA推导的蛋白质序列进行检查发现有许多七肽重复序列,这可能允许通过逐步缺失和可变剪接来改变长度。这些重复序列表明存在α - 螺旋卷曲螺旋结构,它可能在二聚体亚基之间形成相互作用,或者与细胞骨架形成相互作用,或者两者都有。