MOLECULAR CLONING & REGULATION OF HUMAN INDUCIBLE NITRIC OXIDE SYNTHASE GENE

分子克隆

• 批准号: 6221089
• 负责人: DAVID A GELLER
• 金额: $ 0.13万
• 依托单位: MELLON PITTS CORPORATION (MPC CORP)
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6221089
• 关键词: biological products; biomedical resource; computers

We are interested in finding the amino acids and structural features which determine water permeability in the aquaporin (AQP) family of proteins and in the larger family of membrane intrinsic proteins (MIPs). We have expressed the AQP1 protein in yeast secretory vesicles and have also reconstituted yeast-expressed AQP1 into lipid vesicles. We have shown that the protein acts as a water channel in both environments. We are in the process of expressing AQP2 and AQP3 in the same yeast system. We have expressed two closely related proteins (FPS and GlpF) from the MIP family which, although homologous in sequence, do not act as water channels. We would like to find specific amino acids or protein domains which differ among the water channels and non-water channels and which may be important for determining water permeability. To do this, we plan to analyze the sequences and secondary structures of all of the MIP and AQP proteins and look for regions which are conserved or variable. Sites which potentially determine water permeability can then be evaluated experimentally using our yeast expression system. Sequence alignments and secondary structure predictions are possible with the CGC software package which is available from the Pittsburgh Supercomputing Center.

我们感兴趣的是找到氨基酸和结构 决定水通道蛋白（AQP）透水性的特征 在蛋白质家族和膜内 蛋白质（MIPs）。 我们在酵母中表达了AQP1蛋白 分泌囊泡，也重建了酵母表达的AQP1 变成脂质囊泡。 我们已经证明蛋白质就像水一样 两种环境下的渠道。 我们正在表达 AQP2和AQP3在同一酵母系统中。 我们已经表达了两个密切 来自MIP家族的相关蛋白（FPS和GlpF），尽管 在序列上同源，不作为水通道。 我们想 为了找到特定的氨基酸或蛋白质结构域， 水通道和非水通道， 测定透水性。 为此，我们计划分析 所有MIP和AQP蛋白的序列和二级结构 寻找保守或可变的区域。 的遗址开展 然后可以评估潜在确定的水渗透性 实验性地使用我们的酵母表达系统。 序列比对 和二级结构预测是可能的与CGC软件 该软件包可从匹兹堡超级计算中心获得。