Schulze-Gahmen U, Prinz H, Glatter U, Beyreuther K
EMBO J. 1985 Jul;4(7):1731-7. doi: 10.1002/j.1460-2075.1985.tb03843.x.
In an attempt to assign secondary structure elements to protein primary structures with antibodies, we synthesized a model peptide (beta-peptide: TVTVTDPGQTVTY) with a putative beta-turn structure and analysed the anti-peptide antibodies for their specificity towards the turn sequence. At least 50% of the peptide fraction adopts the intended conformation of a beta-turn (DPGQ) inserted between the two segments of an antiparallel beta-sheet structure. The specific anti-beta-peptide antibodies of the hyperimmune response bind the beta-turn containing epitope of the immunogenic beta-peptide with a three orders of magnitude higher affinity than the synthetic control peptide (Gly-peptide: GGGGGDPGQGGGG). The affinity of the antibodies with specificity for the beta-turn region increases from the primary to the hyperimmune response. Therefore, probing of secondary structure elements, i.e., of individual beta-turn regions, by anti-peptide antibodies now seems feasible for proteins of known sequence and may result in sequence assignments of secondary structures.
为了用抗体将二级结构元件赋予蛋白质一级结构,我们合成了一种具有假定β-转角结构的模型肽(β-肽:TVTVTDPGQTVTY),并分析了抗肽抗体对转角序列的特异性。至少50%的肽段采用了插入反平行β-折叠结构两段之间的预期β-转角(DPGQ)构象。超免疫反应产生的特异性抗β-肽抗体与免疫原性β-肽含β-转角的表位结合,其亲和力比合成对照肽(甘氨酸肽:GGGGGDPGQGGGG)高三个数量级。对β-转角区域具有特异性的抗体的亲和力从初次免疫反应到超免疫反应逐渐增加。因此,现在看来,用抗肽抗体探测二级结构元件,即单个β-转角区域,对于已知序列的蛋白质是可行的,并且可能导致二级结构的序列归属。