Sterk P, Booij H, Schellekens G A, Van Kammen A, De Vries S C
Department of Molecular Biology, Agricultural University Wageningen, The Netherlands.
Plant Cell. 1991 Sep;3(9):907-21. doi: 10.1105/tpc.3.9.907.
A cDNA corresponding to a 10-kD protein, designated extracellular protein 2 (EP2), that is secreted by embryogenic cell cultures of carrot was obtained by expression screening. The derived protein sequence and antisera against heterologous plant lipid transfer proteins identified the EP2 protein as a lipid transfer protein. Protein gel blot analysis showed that the EP2 protein is present in cell walls and conditioned medium of cell cultures. RNA gel blot analysis revealed that the EP2 gene is expressed in embryogenic cell cultures, the shoot apex of seedlings, developing flowers, and maturing seeds. In situ hybridization showed expression of the EP2 gene in protoderm cells of somatic and zygotic embryos and transient expression in epidermis cells of leaf primordia and all flower organs. In the shoot apical meristem, expression is found in the tunica and lateral zone. In maturing seeds, the EP2 gene is expressed in the outer epidermis of the integument, the seed coat, and the pericarp epidermis, as well as transiently in between both mericarps. Based on the extracellular location of the EP2 protein and the expression pattern of the encoding gene, we propose a role for plant lipid transfer proteins in the transport of cutin monomers through the extracellular matrix to sites of cutin synthesis.
通过表达筛选获得了一个与10 kD蛋白对应的cDNA,该蛋白命名为细胞外蛋白2(EP2),由胡萝卜胚性细胞培养物分泌。推导的蛋白序列以及针对异源植物脂质转移蛋白的抗血清将EP2蛋白鉴定为脂质转移蛋白。蛋白质凝胶印迹分析表明,EP2蛋白存在于细胞培养物的细胞壁和条件培养基中。RNA凝胶印迹分析显示,EP2基因在胚性细胞培养物、幼苗的茎尖、发育中的花和成熟种子中表达。原位杂交显示,EP2基因在体细胞胚和合子胚的原表皮细胞中表达,并在叶原基和所有花器官的表皮细胞中瞬时表达。在茎尖分生组织中,在 tunica和侧区发现表达。在成熟种子中,EP2基因在珠被的外表皮、种皮和果皮表皮中表达,并在两个分果之间瞬时表达。基于EP2蛋白的细胞外定位和编码基因的表达模式,我们提出植物脂质转移蛋白在角质单体通过细胞外基质运输到角质合成部位中发挥作用。