Tresguerres Martin, Salmerón Cristina
Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, United States.
Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, United States.
Methods Enzymol. 2018;605:525-549. doi: 10.1016/bs.mie.2018.02.022. Epub 2018 Apr 11.
The enzyme soluble adenylyl cyclase (sAC) is the most recently identified source of the messenger molecule cyclic adenosine monophosphate. sAC is evolutionarily conserved from cyanobacteria to human, is directly stimulated by [Formula: see text] ions, and can act as a sensor of environmental and metabolic CO, pH, and [Formula: see text] levels. sAC genes tend to have multiple alternative promoters, undergo extensive alternative splicing, be translated into low mRNA levels, and the numerous sAC protein isoforms may be present in various subcellular localizations. In aquatic organisms, sAC has been shown to mediate various functions including intracellular pH regulation in coral, blood acid/base regulation in shark, heart beat rate in hagfish, and NaCl absorption in fish intestine. Furthermore, sAC is present in multiple other species and tissues, and sAC protein and enzymatic activity have been reported in the cytoplasm, the nucleus, and other subcellular compartments, suggesting even more diverse physiological roles. Although the methods and experimental tools used to study sAC are conventional, the complexity of sAC genes and proteins requires special considerations that are discussed in this chapter.
可溶性腺苷酸环化酶(sAC)是信使分子环磷酸腺苷的最新发现来源。sAC在从蓝细菌到人类的进化过程中保守存在,直接受[公式:见正文]离子刺激,可作为环境和代谢中二氧化碳、pH值及[公式:见正文]水平的传感器。sAC基因往往有多个可变启动子,经历广泛的可变剪接,转录成低水平的mRNA,众多sAC蛋白异构体可能存在于不同的亚细胞定位中。在水生生物中,sAC已被证明可介导多种功能,包括珊瑚细胞内pH调节、鲨鱼血液酸碱调节、盲鳗心率调节以及鱼肠中氯化钠吸收。此外,sAC存在于多种其他物种和组织中,在细胞质、细胞核及其他亚细胞区室中均有sAC蛋白和酶活性的报道,提示其生理作用更为多样。尽管用于研究sAC的方法和实验工具是常规的,但sAC基因和蛋白质的复杂性需要特殊考虑,本章将对此进行讨论。