Max-Plank Institute for Plant Breeding Research, Cologne, Germany.
Plant Cell Rep. 2009 Nov;28(11):1747-58. doi: 10.1007/s00299-009-0775-2. Epub 2009 Oct 2.
Whereas the important plant growth regulator auxin has multiple effects in flowering plants, it induces a specific cell differentiation step in the filamentous moss protonema. Here, we analyse the presence of classical auxin-binding protein (ABP1) homologues in the moss Funaria hygrometrica. Microsomal membranes isolated from protonemata of F. hygrometrica have specific indole acetic acid-binding sites, estimated to be about 3-5 pmol/mg protein with an apparent dissociation constant (K (d)) between 3 and 5 microM. Western analyses with anti-ABP1 antiserum detected the canonical endoplasmic reticulum (ER)-localised 22-24 kDa ABP1 in Zea mays, but not in F. hygrometrica. Instead, polypeptides of 31-33 and 46 kDa were labelled in the moss as well as in maize. In F. hygrometrica these proteins were found exclusively in microsomal membrane fractions and were confirmed as ABPs by photo-affinity labelling with 5-azido-[7-(3)H]-indole-3-acetic acid. Unlike the classical corn ABP1, these moss ABPs did not contain the KDEL ER retention sequence. Consistently, the fully sequenced genome of the moss Physcomitrella patens, a close relative of F. hygrometrica, encodes an ABP1-homologue without KDEL sequence. Our study suggests the presence of putative ABPs in F. hygrometrica that share immunological epitopes with ABP1 and bind auxin but are different from the classical corn ABP1.
尽管重要的植物生长调节剂生长素在开花植物中有多种作用,但它在丝状藓原丝体中诱导特定的细胞分化步骤。在这里,我们分析了藓 Funaria hygrometrica 中是否存在经典的生长素结合蛋白 (ABP1) 同源物。从 F. hygrometrica 原丝体分离的微粒体膜具有特定的吲哚乙酸结合位点,估计约为 3-5 pmol/mg 蛋白,表观解离常数 (K (d)) 在 3 和 5 microM 之间。用抗 ABP1 抗血清进行的 Western 分析检测到玉米 Zea mays 中经典的内质网 (ER) 定位的 22-24 kDa ABP1,但在 F. hygrometrica 中未检测到。相反,在藓和玉米中均标记了 31-33 和 46 kDa 的多肽。在 F. hygrometrica 中,这些蛋白质仅存在于微粒体膜部分中,并通过与 5-叠氮-[7-(3)H]-吲哚-3-乙酸的光亲和标记被确认为 ABPs。与经典的玉米 ABP1 不同,这些藓 ABPs 不包含 KDEL ER 保留序列。一致地,藓 Physcomitrella patens 的全序列基因组,它是 F. hygrometrica 的近亲,编码了一个没有 KDEL 序列的 ABP1 同源物。我们的研究表明,在 F. hygrometrica 中存在与 ABP1 具有免疫表位且结合生长素的假定 ABPs,但与经典的玉米 ABP1 不同。