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A Cysteine-Rich Extracellular Protein LAT52 Interacts with the Extracellular Domain of the Pollen Receptor Kinase LePRK2

机译:半胱氨酸丰富的细胞外蛋白LAT52与花粉受体激酶LePRK2的胞外域相互作用。

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摘要

Pollen germination and pollen tube growth are thought to require extracellular cues, but how these cues are perceived and transduced remains largely unknown. Pollen receptor kinases are plausible candidates for this role; they might bind extracellular ligands and thereby mediate cytoplasmic events required for pollen germination and pollen tube growth. To search for pollen-expressed ligands for pollen receptor kinases, we used the extracellular domains of three pollen-specific receptor kinases of tomato (LePRK1, LePRK2, and LePRK3) as baits in a yeast two-hybrid screen. We identified numerous secreted or plasma membrane–bound candidate ligands. One of these, the Cys-rich protein LAT52, was known to be essential during pollen hydration and pollen tube growth. We used in vivo coimmunoprecipitation to demonstrate that LAT52 was capable of forming a complex with LePRK2 in pollen and to show that the extracellular domain of LePRK2 was sufficient for the interaction. Soluble LAT52 can exist in differently sized forms, but only the larger form can interact with LePRK2. We propose that LAT52 might be a ligand for LePRK2.
机译:花粉萌发和花粉管生长被认为需要细胞外线索,但是如何感知和转导这些线索仍然未知。花粉受体激酶可能是这种作用的候选者。它们可能结合细胞外配体,从而介导花粉萌发和花粉管生长所需的细胞质事件。为了搜索花粉受体激酶的花粉表达配体,我们在酵母两杂交筛选中使用了番茄的三个花粉特异性受体激酶(LePRK1,LePRK2和LePRK3)的胞外域作为诱饵。我们鉴定了许多分泌或质膜结合的候选配体。其中之一,富含Cys的蛋白LAT52,在花粉水合作用和花粉管生长过程中必不可少。我们使用了体内共免疫沉淀技术来证明LAT52能够与花粉中的LePRK2形成复合物,并表明LePRK2的胞外域足以进行相互作用。可溶性LAT52可以以不同大小的形式存在,但只有较大的形式才能与LePRK2相互作用。我们建议,LAT52可能是LePRK2的配体。

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