Stevenson-Paulik Jill, Chiou Shean-Tai, Frederick Josh P, dela Cruz June, Seeds Andrew M, Otto James C, York John D
Department of Pharmacology and Cancer Biology, Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
Methods. 2006 Jun;39(2):112-21. doi: 10.1016/j.ymeth.2006.05.012.
Recent discoveries that provide a link between inositol phosphate (IP) signaling and fundamental cellular processes evoke many exciting new hypotheses about IP function, and underscore the importance of understanding how IP synthesis is regulated. Central to studies of IP metabolism is the essential development of efficient, fast, and reproducible methods for quantitative analysis of IPs in systems ranging from simple cell cultures to more complex tissues and whole organisms. Additionally, in many cases there is a need to pharmacologically and/or genetically alter IP kinase and phosphatase activities in order to visualize low abundance inositol signaling messengers. Here, we describe updated methods for rapid analysis of IP metabolism in normal and genetically manipulated Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Mus musculus, and Homo sapiens.
近期的发现揭示了肌醇磷酸(IP)信号传导与基本细胞过程之间的联系,引发了许多关于IP功能的令人兴奋的新假说,并强调了理解IP合成调控方式的重要性。IP代谢研究的核心是开发高效、快速且可重复的方法,用于在从简单细胞培养到更复杂组织及整个生物体的系统中对IP进行定量分析。此外,在许多情况下,需要通过药理学和/或遗传学手段改变IP激酶和磷酸酶的活性,以便观察低丰度的肌醇信号信使。在此,我们描述了用于快速分析正常及基因操作的酿酒酵母、拟南芥、黑腹果蝇、小家鼠和人类中IP代谢的更新方法。