Higgins J A, Barrnett R J
J Cell Biol. 1971 Jul;50(1):102-20. doi: 10.1083/jcb.50.1.102.
A study of the fine structural localization of the acyltransferases of the monoglyceride and alpha-glycerophosphate pathways for triglyceride synthesis in the intestinal absorptive cell is reported. Glutaraldehyde-fixed tissue was found to synthesize diglyceride and triglyceride from monopalmitin and palmityl CoA, and parallel morphological studies showed the appearance of lipid droplets in the smooth endoplasmic reticulum of the absorptive cell. Glutaraldehyde-fixed tissue also synthesized triglyceride from alpha-glycerophosphate, although this enzyme system was more susceptible to fixation than the monoglyceride pathway acyltransferases. Cytochemical methods for the localization of free CoA were based (a) on the formation of the insoluble lanthanium mercaptide of CoA and (b) on the reduction of ferricyanide by CoA to yield ferrocyanide which forms an insoluble precipitate with manganous ions. By these methods the monoglyceride pathway acyltransferases were found to be located mainly on the inner surface of the smooth endoplasmic reticulum. The alpha-glycerophosphate pathway acyltransferases were localized mainly on the rough endoplasmic reticulum. Activity limited to the outer cisternae of the Golgi membranes occurred with both pathways. The possible organization of triglyceride absorption and chylomicron synthesis is discussed in view of these results.
本文报道了一项关于肠道吸收细胞中甘油三酯合成的甘油单酯和α-甘油磷酸途径的酰基转移酶精细结构定位的研究。研究发现,经戊二醛固定的组织能够利用单棕榈酸甘油酯和棕榈酰辅酶A合成甘油二酯和甘油三酯,同时进行的形态学研究显示吸收细胞的滑面内质网中出现了脂滴。经戊二醛固定的组织也能利用α-甘油磷酸合成甘油三酯,不过该酶系统比甘油单酯途径的酰基转移酶更容易受到固定的影响。用于定位游离辅酶A的细胞化学方法基于:(a)辅酶A的不溶性硫醇镧的形成;(b)辅酶A将铁氰化物还原生成亚铁氰化物,亚铁氰化物与锰离子形成不溶性沉淀。通过这些方法发现,甘油单酯途径的酰基转移酶主要位于滑面内质网的内表面。α-甘油磷酸途径的酰基转移酶主要定位于粗面内质网。两种途径在高尔基体膜的外侧潴泡均有活性。鉴于这些结果,本文讨论了甘油三酯吸收和乳糜微粒合成的可能组织方式。