Characterization of the Digitalis Receptor and Digitalis Mimics

;​我不知道，我不知道，我不知道。l0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 02 3 4 5 6 7 8 9:；F Microsoft Word 6.0文档MSWordDoc。6;​E + E @t E u ~ u E + E = t F F E u V t E u n E u - F V V RP X # t0 E + Hu& 9 E u 9 u u W O $ ^_ v ~ F V ~ ~ t e tq t&amp ; ~ '@;E ~#} u E @；本研究的长期目标是了解阳离子转运体Na，K atp酶的结构和功能，特别感兴趣的是阐明洋地黄（心脏糖苷）结合和抑制该酶的分子机制。P.I.建议使用一种相对较新的，也许是非常规的方法来鉴定和表征可能模仿糖苷的短肽序列，糖苷是基于非肽类固醇的植物产物。这些肽将从含有“随机肽库”的噬菌体中选择。这些选择的多肽的多样性将帮助我们了解糖苷是如何结合的，以及它们的作用是如何被修饰的。此外，由于酶的洋地黄结合位点尚未确定，因此P.I.还将尝试使用抗独特型抗体（提升为抗地高辛抗体）来识别结合位点。随机肽库也将用于确定是否有可能生成模型洋地黄结合位点。这项工作有可能使我们能够识别作为天然调节蛋白的血清蛋白。Na， k -ATP酶是一种膜结合酶，它利用ATP形式的能量在细胞膜上运输钠离子和钾离子。正是这两种重要阳离子水平的维持，推动了氨基酸、葡萄糖、磷酸盐和钙等小分子的主动运输，并为神经和肌肉细胞的动作电位提供了基础。Na， k - atp酶也是洋地黄的受体，洋地黄是一种植物糖苷，它以一种高度特异性的方式抑制该酶。该项目使用一种新的免疫学方法来了解更多关于洋地黄结合的机制，并且应该有助于确定长期寻求的天然或内源性调节酶. ....的分子()()))()() ;Oh +' 0 $ H l D H R:\WWUSER\TEMPLATE\NORMAL。DOT marcia steinberg marcia steinberg @ ？n @ @ (1) n @ (1) n @ (1) n @ (1) n @ (1) n @ (2) n @ (1@ NZ微软Word 6.0e 3 e " " " " " " " L L L L L L d n L C x | | | | | | | _ P T 4“ | | | | | | ” “ | x | | | | ” | " | 6 " " " " | | 3 | 9422022球本研究的长期目标是获得对结构和阳离子转运体Na，K atp酶的功能，特别感兴趣的是阐明洋地黄（心脏糖苷）结合和抑制该酶的分子机制。P.I.建议使用一种相对较新的，也许是非常规的方法来鉴定和表征可能模仿糖苷的短肽序列，糖苷是基于非肽类固醇的植物产物。这些多肽将从含有“随机p”的噬菌体中选择

; R o o t E n t r y F ! Lr C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l mÕGr mÕGr 2 3 4 5 6 7 8 9 : ; F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; E +E @t E u ~ u E +E = u t F E U F V t E U v n E U - F V v RP X # t0 E + Hu& 9E u 9U u W u u O$ ^_ v ~ F V ~ ~ t e tq t& ~ '@;E ~# } u E @;E | } } 9422022 Ball The long term goal of this research is to gain an understanding of the structure and functioning of the cation transporter, Na,K ATPase, with a particular interest in elucidating the molecular mechanisms of digitalis (cardiac glycosides) binding and inhibition of this enzyme. The P.I. proposes to use a relatively new and perhaps unconventional approach to identify and characterize short peptide sequences which may mimic the glycosides, which are nonpeptidic steroid based plant products. The peptides will be selected from bacteriophage contained "random peptide libraries". The diversity of these selected peptides will help us understand how the glycosides bind and how their actions may be modified. In addition, since the enzyme's digitalis binding site has not been determined, the P.I. will also attempt to use anti idiotypic antibodies (raised to anti digoxin antibodies) to identify the binding site. The random peptide libraries will also be used to determine whether it is possible to generate a model digitalis binding site(s) This work has the potential of enabling us to identify serum proteins which serve as natural regulatory proteins. %%% The Na,K-ATPase is a membrane bound enzyme that uses energy in the form of ATP to transport sodium and potassium ions across the cell membrane. It is this maintenance of the levels of these two important cations that is the driving force for the active transport of small molecules such as amino acids, glucose, phosphate and calcium, as well as providing the basis for the action potential in nerve and muscle cells. The Na, K-ATPase is also the receptor for digitalis, a plant glycoside, which inhibits the enzyme in a highly specific manner. This project uses a new immunological approach to learn more about the mechanism of digitalis binding, and should in addition help in identifying the long sought after natural, or endogenous, molecule that acts to regulate the enzyme. ....()()))()() ; Oh +' 0 $ H l D h R:\WWUSER\TEMPLATE\NORMAL.DOT marcia steinberg marcia steinberg @ ? n @ @ (r S u m m a r y I n f o r m a t i o n ( 1 @ NZ Microsoft Word 6.0 2 e 3 e " " " " " " " L L L L L d n L C x | | | | | | | _ P T 4 " | | | | | | " " | x | | | | " | " | 6 " " " " | | 3 | 9422022 Ball The long term goal of this research is to gain an understanding of the structure and functioning of the cation transporter, Na,K ATPase, with a particular interest in elucidating the molecular mechanisms of digitalis (cardiac glycosides) binding and inhibition of this enzyme. The P.I. proposes to use a relatively new and perhaps unconventional approach to identify and characterize short peptide sequences which may mimic the glycosides, which are nonpeptidic steroid based plant products. The peptides will be selected from bacteriophage contained "random p