Nucleosidediphosphatase activity in the Golgi apparatus and its usefulness for cytological studies.
Nucleosidediphosphatase activity in the Golgi apparatus and its usefulness for cytological studies.
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高尔基体中核苷二磷酸酶的活性及其在细胞学研究中的用途。
DOI：
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发表时间：
1961
影响因子：
11.1
通讯作者：
S. Goldfischer
中科院分区：
文献类型：
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作者：
A. B. Novikoff;S. Goldfischer
胞苷和腺苷;焦磷酸硫胺素（Sigma）;磷酸核黄素（Sigma）;和无机焦磷酸盐（Merck）。这些化合物中的最后一种在Naidoo和Pratt的系统中进行了测试，用锰取代镁离子。还用二磷酸吡啶核苷酸（DPN）、黄素腺嘌呤二核苷酸（FAD）、鸟苷二磷酸甘露糖（GdG）和尿苷二磷酸葡萄糖（UDPG）进行了初步实验。使用这些底物，将3-5 mg酸性磷酸酶[小麦胚芽（Sigma）]或碱性磷酸酶[小牛肠（Worthington）]添加到孵育培养基中，以从焦磷酸盐的任何水解产生的单磷酸盐中释放磷酸盐残留物。通过酶键。研究了钙、镁和锰添加物的影响，以及培养基pH值的变化。TritonX-100的作用是通过在0 ℃和室温下用~ 0.25%的蒸馏水溶液处理切片5-60分钟，用几次冷蒸馏水的更换来升高它们以除去Triton，然后孵育来研究的。将甲醛固定的大鼠附睾、大鼠脑（大脑）和小鼠Friend白血病的组织块或冷冻切片孵育，然后用四氧化锇处理，并以通常的方式进行处理，用于电子显微镜检查。这些实验是由爱德华·德雷纳博士进行的，将在别处报道。除洋葱根、根尖外，所有组织均采用Aoyama银法制作石蜡切片，以显示高尔基体。
cytidine, and adenosine; thiamine pyrophosphate (Sigma); riboflavin phosphate (Sigma); and inorganic pyrophosphate (Merck). The last of these compounds was tested in the system of Naidoo and Pratt,"' with manganese substituted for magnesium ions. Preliminary experiments were also done with diphosphopyridine nucleotide (DPN), flavin adenine dinucleotide (FAD), guanosine diphosphate mannose (GDPM), and uridine diphosphate glucose (UDPG). With these substrates, 3-5 mg of either acid phosphatase [wheat germ (Sigma)] or alkaline phosphatase [calf intestine (Worthington)] was added to the incubation medium, in order to liberate the phosphate residues from monophosphates resulting from any hydrolysis of the pyrophosphate bonds by the enzyme. The effects of calcium, magnesium, and manganese additions were studied, as were variations :in the pH of the medium. The effects of Triton X-100 were studied by treating sections with -0.25% solutions in distilled water for 5-60 minutes at 00C and room temperature, rising them with .several changes of cold distilled water to remove the Triton, and then incubating. Tissue blocks or frozen sections of formaldehyde-fixed rat epididymis, rat brain (cerebrum), and the mouse Friend leukemia were incubated and then treated with osmium tetroxide and processed, in the usual manner, for electron microscopy. These experiments, performed by Dr. Edward Essner, will be reported elsewhere. Paraffin sections were made of all tissues except the onion root, tips, following the Aoyama silver method for demonstrating the Golgi apparatus.12