SGER: Development and Testing of a "Generic" K+ Channel Antibody

SGER：“通用”K 通道抗体的开发和测试

• 批准号: 9320336
• 负责人: Gerald Berkowitz
• 金额: $ 5万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9320336&HistoricalAwards=false
• 关键词: SGER; Development; Testing; Generic; K+

9320336 Berkowitz, Gerald A. Progress toward the molecular characterization of K+-channels is hindered by lack of a probe which can selectively identify these proteins in membrane preparations, and in translation products of cDNA's. A research program was initiated and aimed at developing such a probe by generating antibodies to a short peptide corresponding to the deduced amino acid sequence in the pore of the protein which acts as a selectivity filter, and is highly conserved in known K+ channels. The objective of the work is to evaluate the usefulness of the antibody by examining it's affinity for a broad range of K+ channels, and it's selectivity for this class of proteins. The work will involve immunoblot analysis of the antibody's reactivity with known K+ channels in native membranes (chloroplast inner envelope) and in heterologous systems expressing products of K+ channel cDNA's (KAT1 K+ channel cDNA in yeast, drkl cDNA in kidney cell lines, and Shab, Shaw, Shal and KAT1 cDNAs in E. coli). %%% The laboratory made use of an unusual technique to raise a specific antibody to a specific part of the K+ channel protein that is common to all known K+ channels. The goal of the project is to show proof that this antibody is "generic", that means that it recognizes all K+ channels, irrespective of the system it is used in. Since it is presently generally assumed that the method used in the antibody production will not work, and therefore no specific probes can be generated, such a proof is necessary. This is a "high risk, high reward" pioneering study that is well suited to the Small Grants for Exploratory R esearch program. *** } ! ! ! F ( Times New Roman Symbol & Arial " h " $" 5 Dr. Geza Hrazdina Dr. Geza Hrazdina

9320336 Berkowitz，Gerald A. K+通道的分子表征的进展由于缺乏可以选择性地鉴定膜制备物和cDNA的翻译产物中的这些蛋白质的探针而受到阻碍。 启动了一项研究计划，旨在通过产生与蛋白质孔中推导的氨基酸序列对应的短肽的抗体来开发这种探针，该蛋白质孔用作选择性过滤器，并且在已知的K+通道中高度保守。 这项工作的目的是通过检查抗体对广泛的K+通道的亲和力以及对这类蛋白质的选择性来评估抗体的有用性。 这项工作将涉及抗体与天然膜（叶绿体内被膜）和表达K+通道cDNA产物（酵母中的KAT 1 K+通道cDNA、肾细胞系中的drk1 cDNA和大肠杆菌中的Shab、Shaw、Shal和KAT 1 cDNA）的异源系统中已知K+通道的反应性的免疫印迹分析。大肠杆菌）。 该实验室利用一种不寻常的技术，针对所有已知K+通道共有的K+通道蛋白的特定部分，产生了一种特异性抗体。 该项目的目标是证明这种抗体是“通用的”，这意味着它可以识别所有的K+通道，而不管它用于什么系统。 由于目前通常认为抗体生产中使用的方法不起作用，因此不能产生特异性探针，因此需要这样的证明。 这是一项“高风险，高回报”的开创性研究，非常适合探索性研究计划的小额赠款。 *** } ! ! ! F ( Times New Roman 符号Arial " H " $" 5 Geza Hrazdina博士