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使用两性离子去污剂在不使非组蛋白染色体蛋白质变性的情况下进行定量增溶。

Quantitative solubilization of nonhistone chromosomal proteins without denaturation using zwitterionic detergents.

作者信息

Matuo Y, Matsui S, Nishi N, Wada F, Sandberg A A

出版信息

Anal Biochem. 1985 Nov 1;150(2):337-44. doi: 10.1016/0003-2697(85)90520-2.

Abstract

Of three kinds of commercial zwitterionic detergents [SB 12, SB 14, and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps)], SB 12 and Chaps were more useful than SB 14 because of high solubility and less interference with protein assay. Efficiency for protein solubilization at pH 6-9 was higher for SB 12 than for Chaps with either calf thymus chromatin or rat liver nuclei. At pH 9 and ionic strength (I) = 0.35, 1% SB 12 and 1% Chaps were capable of solubilizing about 70% and about 47% of total proteins in rat liver nuclei, respectively. Core histones in rat liver nuclei were extracted to a lesser extent with Chaps than with SB 12. DNA-dependent RNA polymerase and isopeptidase activities were barely inactivated by 1% Chaps at pH 8-9, but isopeptidase activity was inhibited by 0.3% SB 12. These facts indicate that whereas SB 12 is effective for solubilization of whole nuclear proteins, Chaps is suitable for the selective extraction of nonhistone chromosomal proteins without denaturation.

摘要

在三种商业两性离子去污剂[SB 12、SB 14和3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐(Chaps)]中,SB 12和Chaps比SB 14更有用,因为它们具有高溶解性且对蛋白质测定的干扰较小。对于小牛胸腺染色质或大鼠肝细胞核,在pH 6-9时,SB 12溶解蛋白质的效率高于Chaps。在pH 9和离子强度(I)=0.35时,1%的SB 12和1%的Chaps分别能够溶解大鼠肝细胞核中约70%和约47%的总蛋白质。与SB 12相比,Chaps对大鼠肝细胞核核心组蛋白的提取程度较低。在pH 8-9时,1%的Chaps几乎不会使DNA依赖性RNA聚合酶和异肽酶活性失活,但0.3%的SB 12会抑制异肽酶活性。这些事实表明,虽然SB 12对溶解全核蛋白质有效,但Chaps适合于在不发生变性的情况下选择性提取非组蛋白染色体蛋白质。

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