Molecular Characterization of Vacuolar Membrane, Calcium Binding Proteins

液泡功能对调节胞质钙水平至关重要。钙结合蛋白独特的液泡膜已被确定。该项目的直接目标是纯化和表征这些蛋白质的亲和力，容量和阳离子选择性。多克隆抗体将用于研究这些蛋白的组织特异性表达模式，特别是关于液泡化过程和液泡膜系统的生物发生。抗体将进一步用于鉴定编码这些蛋白的表达文库中的cDNA克隆。植物细胞含有大的膜封闭的隔室，它们为植物细胞提供许多生化和结构功能。其中一些是相对“普遍”的功能，通常与溶酶体型室有关；另一些则是植物特有的。因此，液泡在生长发育中起着至关重要的作用。值得注意的是，很少有空泡膜的活性被研究，在分子水平上被鉴定和研究的更少。目前还不清楚这种细胞器的起源，也不清楚液泡膜蛋白是如何不断补充的。这项研究将有助于了解液泡在赋予高等植物对环境的响应能力和进行正常生长发育过程中的作用的分子基础。

Vacuolar function is essential to the regulation of cytosolic calcium levels. Calcium-binding proteins unique to the vacuolar membrane have been identified. The immediate goal of this project is to purify and characterize these proteins in terms of affinity, capacity, and cation selectivity. Polyclonal antibodies will be used to study the tissue-specific expression patterns of these proteins, particularly with regard to the process of vacuolation and biogenesis of the vacuole membrane system. Antibody will further serve to identify cDNA clones in expression libraries encoding these proteins. %%% Plant cells contain large membrane-enclosed compartments, which serve a number of biochemical and structural functions for the plant cell. Some of these are relatively "universal" functions associated generally with lysosome-type compartments; others are unique for plants. Accordingly, the vacuole has an essential role in growth and development. It is remarkable that few vacuolar membrane activities have been examined and fewer still have been identified and examined at the molecular level. There is not yet a clear understanding of either the origins of this organelle or the means by which vacuolar membrane proteins are continuously replenished. This study will contribute to an understanding of the molecular basis for the role of the vacuole in conferring to higher plants the ability to respond to its environment and carry out the normal processes of growth and development.