Hemin inhibits ubiquitin-dependent proteolysis in both a higher plant and yeast.
Hemin inhibits ubiquitin-dependent proteolysis in both a higher plant and yeast.
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氯高铁血红素抑制高等植物和酵母中泛素依赖性蛋白水解作用。
DOI：
10.1021/bi00409a025
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Sullivan,ML
中科院分区：
文献类型：
--
作者：
Vierstra,RD;Sullivan,ML
材料和方法试剂。根据 Haas 和 Wilkinson (1985) 的方法从红细胞中纯化人泛素。蛋清溶菌酶获自 Sigma，并按照先前所述进行纯化（Hough & Rechsteiner，1986）。使用购自 Amersham 的无载体 Na125I (5.6 X 106 Bq//ig)，通过氯胺 T 法（Ciechanover 等，1980）对泛素和溶菌酶进行 125I 放射性标记。 125I-泛素制剂的初始比放射性为(3.9-14) X 103 cpm/pmol，125I-溶菌酶制剂的初始比放射性为(1.6-3.1) X 104 cpm/pmol。氯高铁血红素、原卟啉 IX、ATP、亮肽素、己糖激酶和磷酸肌酸激酶购自 Sigma。中血红素是来自犹他州洛根市卟啉产品公司的产品。将血红素和中血红素的储备溶液 (1 mM) 溶解在 50 mM 中
Materials and MethodsReagents. Human ubiquitin was purified from erythrocytes according to the method of Haas and Wilkinson (1985). Egg white lysozyme was obtained from Sigma and purified as previously described (Hough & Rechsteiner, 1986). Both ubiquitin and lysozyme were radiolabeled with 125I by the chloramine T method (Ciechanover et al., 1980) using car-rier-free Na125I (5.6 X 106 Bq//ig) purchased from Amersham. The initial specific radioactivities for 125I-ubiquitin preparations were (3.9-14) X 103 cpm/pmol, and those for 125I-lysozyme preparations were(1.6-3.1) X 104 cpm/pmol. Hemin, pro-toporphyrin IX, ATP, leupeptin, hexokinase, and phosphocreatine kinase were purchased from Sigma. Mesohemin was a product of Porphyrin Products, Logan, UT. Stock solutions (1 mM) of hemin and mesohemin were dissolved in 50 mM