Molecular and physiological characterisation of a 14-3-3 protein from lily pollen grains regulating the activity of the plasma membrane H+ ATPase during pollen grain germination and tube growth
Molecular and physiological characterisation of a 14-3-3 protein from lily pollen grains regulating the activity of the plasma membrane H+ ATPase during pollen grain germination and tube growth
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百合花粉粒 14-3-3 蛋白在花粉粒萌发和管生长过程中调节质膜 H ATP 酶活性的分子和生理学特征
DOI：
10.1007/s004250000483
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发表时间：
2001
期刊：
影响因子：
4.3
通讯作者：
G. Obermeyer
中科院分区：
文献类型：
--
作者：
H. Pertl;M. Himly;R. Gehwolf;Rosa Kriechbaumer;D. Strasser;W. Michalke;K. Richter;F. Ferreira;G. Obermeyer
抽象的。从麝香百合成熟花粉cDNA文库中克隆了一个14-3-3蛋白。对14-3-3蛋白高度特异的单克隆抗体（MUP 5或MUP 15）识别许多不同百合组织（叶、鳞茎、茎、花药丝、花粉粒、柱头）和其它植物（拟南芥幼苗、大麦重组体14-3-3）的细胞质部分中的30-kDa蛋白。此外，14-3-3蛋白质中检测到的微粒体部分分离的花粉粒和管，和膜结合的14-3-3蛋白质的量以及质膜（PM）H+ ATP酶的量增加，在萌发的花粉粒和管的生长。在细胞质组分中未观察到变化。当花粉粒在含有1 μM梭孢菌素（FC）的萌发培养基中孵育时，微粒体组分中14-3-3蛋白的数量进一步增加，而细胞质组分中14-3-3s的数量减少。Fusicoccin也保护膜结合的14-3-3蛋白在用离液盐KI洗涤后免于解离。此外，FC还能促进百合花粉质膜H+ ATPase活性、萌发频率和花粉管生长速率，表明14-3-3蛋白通过与PM H+ ATPase活性的相互作用调节百合花粉萌发和花粉管生长。
Abstract. A 14-3-3 protein has been cloned and sequenced from a cDNA library constructed from mRNAs of mature pollen grains of Lilium longiflorum Thunb. Monoclonal antibodies (MUP 5 or MUP 15) highly specific against 14-3-3 proteins recognised a 30-kDa protein in the cytoplasmic fraction of many various lily tissues (leaves, bulbs, stems, anther filaments, pollen grains, stigmas) and in other plants (Arabidopsis seedlings, barley recombinant 14-3-3). In addition, 14-3-3 proteins were detected in a microsomal fraction isolated from pollen grains and tubes, and the amount of membrane-bound 14-3-3 proteins as well as the amount of the plasma membrane (PM) H+ ATPase increased during germination of pollen grains and tube growth. No change was observed in the cytoplasmic fraction. A further increase in the amount of 14-3-3 proteins in the microsomal fraction was observed when pollen grains were incubated in germination medium containing 1 μM fusicoccin (FC) whereas the number of 14-3-3s in the cytoplasmic fraction decreased. Fusicoccin also protected membrane-bound 14-3-3 proteins from dissociation after washing with the chaotropic salt KI. Furthermore, FC stimulated the PM H+ ATPase activity, the germination frequency and the growth rate of pollen tubes, thus indicating that a modulation of the PM H+ ATPase activity by interaction with 14-3-3 proteins may regulate germination and tube growth of lily pollen.