Reinard T, Achmus H, Walther A, Rescher U, Klämbt D, Jacobsen H J
LG Molekulargenetik, Universität Hannover, F.R.G.
Plant Cell Physiol. 1998 Aug;39(8):874-8. doi: 10.1093/oxfordjournals.pcp.a029447.
Several approaches were successfully performed to directly assign and characterize auxin binding of ABP44 in gel. The 44 kDa high affinity auxin binding protein ABP44 from pea was tested for its ability to bind 5-azido-[7-3H]-IAA in photoaffinity labeling experiments. Competition experiments with several auxin analogues confirm data published previously (Reinard and Jacobsen 1995). Critical reflections of the limitations of the method are also discussed. Immunostaining using the antibody D16 (Napier and Venis 1992), which is directed against the putative binding site of ABP1, revealed that ABP44's auxin binding site is at least partially related to the corresponding site of ABP1. Nevertheless, both proteins do not share any further immunological relationships. Our results with D16 recommend a careful reconsideration of data published by other authors. Furthermore, a 80 kDa, dimeric glutathione dependent formaldehyde dehydrogenase (FDH) from mung bean, described recently, was found to be different from ABP44. In contrast to the described FDH, ABP44 exhibited no FDH activity.
已经成功采用了几种方法来直接在凝胶中鉴定和表征ABP44的生长素结合情况。在光亲和标记实验中，对来自豌豆的44 kDa高亲和力生长素结合蛋白ABP44结合5-叠氮基-[7-³H]-IAA的能力进行了测试。用几种生长素类似物进行的竞争实验证实了先前发表的数据（Reinard和Jacobsen，1995年）。还讨论了对该方法局限性的批判性思考。使用针对ABP1假定结合位点的抗体D16（Napier和Venis，1992年）进行免疫染色，结果显示ABP44的生长素结合位点至少部分与ABP1的相应位点相关。然而，这两种蛋白质没有任何其他免疫关系。我们用D16得到的结果建议其他作者仔细重新审视已发表的数据。此外，最近描述的来自绿豆的80 kDa二聚体谷胱甘肽依赖性甲醛脱氢酶（FDH）与ABP44不同。与所描述的FDH相反，ABP44没有表现出FDH活性。
