Sáez D E, Figueroa C D, Concha I I, Slebe J C
Instituto de Bioquímica, Universidad Austral de Chile, Chile.
J Cell Biochem. 1996 Dec 15;63(4):453-62. doi: 10.1002/(SICI)1097-4644(19961215)63:4%3C453::AID-JCB7%3E3.0.CO;2-S.
The localization of fructose 1,6-bisphosphatase (D-Fru-1,6-)2-1-phosphohydrolase, EC 3.1.3.11) in rat kidney and liver was determined immunohistochemically using a polyclonal antibody raised against the enzyme purified from pig kidney. The immunohistochemical analysis revealed that the bisphosphatase was preferentially localized in hepatocytes of the periportal region of the liver and was absent from the perivenous region. Fructose-1,6-bisphosphatase was also preferentially localized in the cortex of the kidney proximal tubules and was absent in the glomeruli, loops of Henle, collecting and distal tubules, and in the renal medulla. As indicated by immunocytochemistry using light microscopy and confirmed with the use of reflection confocal microscopy, the enzyme was preferentially localized in a perinuclear position in the liver and the renal cells. Subcellular fractionation studies followed by enzyme activity assays revealed that a majority of the cellular fructose-1,6-bisphosphatase activity was associated to subcellular particulate structures. Overall, the data support the concept of metabolic zonation in liver as well as in kidney, and establish the concept that the Fructose-1,6-bisphosphatase is a particulate enzyme that can not be considered a soluble enzyme in the classical sense.
利用针对从猪肾中纯化的果糖1,6 -二磷酸酶（D - Fru - 1,6 -）2 - 1 -磷酸水解酶（EC 3.1.3.11）制备的多克隆抗体，通过免疫组织化学方法确定了该酶在大鼠肾脏和肝脏中的定位。免疫组织化学分析显示，双磷酸酶优先定位于肝脏门周区域的肝细胞中，而在肝静脉周围区域不存在。果糖 - 1,6 -二磷酸酶也优先定位于肾近端小管的皮质中，在肾小球、髓袢、集合管和远端小管以及肾髓质中不存在。如通过光学显微镜免疫细胞化学所示并经反射共聚焦显微镜证实，该酶优先定位于肝脏和肾细胞的核周位置。随后进行酶活性测定的亚细胞分级分离研究表明，大部分细胞果糖 - 1,6 -二磷酸酶活性与亚细胞颗粒结构相关。总体而言，这些数据支持肝脏和肾脏中代谢分区的概念，并确立了果糖 - 1,6 -二磷酸酶是一种颗粒酶的概念，不能将其视为传统意义上的可溶性酶。
