Nigam V N, Fridland A
Biochem J. 1967 Nov;105(2):505-13. doi: 10.1042/bj1050505.
Liver homogenates of avian species, but not of mammals, form glycogen from glucose, mannose, fructose and galactose. Incorporation of labelled glucose, fructose and mannose, but not of labelled galactose, into glycogen is diluted isotopically by unlabelled glucose. Except for fructose, glycogen formation from other substrates by pigeon liver homogenates compares favourably with that from the same substrates in pigeon liver slices. Optimum conditions for glycogen synthesis from glucose by pigeon liver homogenate are: medium of incubation, 0.175m-sucrose-45mm-potassium chloride-15mm-glycylglycine buffer, pH7.5; concentration of substrate, 15mm; concentration of tissue, less than 120mg./ml.; temperature of incubation, 37-43 degrees ; atmosphere, oxygen. Uncouplers of oxidative phosphorylation, Ca(2+), EDTA, PP(i), 2-deoxyglucose 6-phosphate and microsomal fraction of rat liver are inhibitory to glycogen synthesis from glucose. Starvation of pigeons for 24 and 48hr. leads to a slight stimulation of glycogen synthesis in their liver homogenates as compared with fed controls. Pigeon liver homogenates can be separated into subcellular fractions that on reconstitution can synthesize glycogen. All the enzymes of the glycogen pathway except soluble high-K(m) glucokinase are present in pigeon liver.
鸟类肝脏匀浆而非哺乳动物肝脏匀浆可利用葡萄糖、甘露糖、果糖和半乳糖合成糖原。标记的葡萄糖、果糖和甘露糖可掺入糖原,但标记的半乳糖则不然,未标记的葡萄糖会使掺入糖原的标记物发生同位素稀释。除果糖外,鸽肝匀浆利用其他底物合成糖原的能力与鸽肝切片利用相同底物合成糖原的能力相当。鸽肝匀浆由葡萄糖合成糖原的最佳条件为:孵育介质为0.175m - 蔗糖 - 45mm - 氯化钾 - 15mm - 甘氨酰甘氨酸缓冲液,pH7.5;底物浓度为15mm;组织浓度小于120mg./ml.;孵育温度为37 - 43度;气体环境为氧气。氧化磷酸化解偶联剂、Ca(2+)、EDTA、PP(i)、2 - 脱氧葡萄糖6 - 磷酸以及大鼠肝脏微粒体组分均抑制由葡萄糖合成糖原。与喂食对照组相比,使鸽子饥饿24小时和48小时会导致其肝脏匀浆中糖原合成略有增加。鸽肝匀浆可分离成亚细胞组分,重新组合后能够合成糖原。糖原合成途径中除可溶性高K(m)葡萄糖激酶外的所有酶均存在于鸽肝中。