A HEME-PEPTIDE AS AN ULTRASTRUCTURAL TRACER
A HEME-PEPTIDE AS AN ULTRASTRUCTURAL TRACER
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血红素肽作为超结构示踪剂
DOI：
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发表时间：
1970
影响因子：
3.2
通讯作者：
N. Feder
中科院分区：
文献类型：
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作者：
N. Feder
具有各种分子量的血红素蛋白，包括过氧化氢酶、髓过氧化物酶、乳过氧化物酶、辣根过氧化物酶（HRP）和细胞色素c，已被用作超微结构示踪剂（参见Karnovsky和Rice. J. Histochem.细胞化学十七点七百五十一分。1969年）。这些血红素蛋白通过它们的过氧化活性在电子显微镜下被证明。其中最小的是细胞色素c（同上），分子量为12，000，但即使是更小的示踪分子也显然可用于研究组织和细胞的渗透性。我已经制备了一种血红素肽（图1），它可以用作超微结构示踪剂和dem（Heinrich Mack公司，通过H. J.詹森公司，新泽西州蒙特克莱尔）在2000 ml水中与160 mb 1 N HCl混合。向该溶液中加入在30 ml 0.1 N HCl中的500 mg胃蛋白酶（三次结晶，Pentex Corporation）。在室温下1天后，用1N NaOH将溶液调至pH 8.5，并向每升溶液中加入610 g固体硫酸铵，使溶液用硫酸铵饱和85%。通过过滤收集红色沉淀物，用85%饱和硫酸铵洗涤并溶解于1000 ml 0.01 N HCl中。过滤溶液并用0.5 N HCl调至pH 1.6。向其中
Hemoproteins with various molecular weights, including catalase, myeloperoxidase, lactoperoxi dase, horseradish peroxidase (HRP) and cytochrome c, have been used as ultrastructural tracers (see Karnovsky and Rice. J. Histochem. Cytochem. 17: 751. 1969). These hemoproteins are demonstrated for electron microscopy through their peroxidatic activity. The smallest of these is cytochrome c (ibid.),with a molecular weight of 12,000, but even smaller tracer molecules would clearly be useful for studying the permeability of tissues and cells. I have prepared a heme-peptide (Fig. 1) that can be used as an ultrastructural tracer and dem(Heinrich Mack Company, via H. J. Jensen, Inc., Montclair, N.J.) in 2000 ml water was mixed with 160mb 1 N HCI. To this solution was added 500 mg pepsin (three times crystallized, Pentex Corporation) in 30 ml 0.1 N HC1. After 1 day at room temperature the solution was brought to pH 8.5 with 1 N NaOH, and 610 g solid ammonium sulfate was added to each liter, making the solution 85% saturated with ammonium sulfate. The red precipitate was collected by filtration, washed with 85% saturated ammonium sulfate and dissolved in 1000 ml 0.01 N HC1. The solution was filtered and brought to pH 1.6 with 0.5 N HC1. To this was