Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate.
Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate.
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在十二烷基硫酸钠存在下，聚丙烯酰胺凝胶电泳后酶的复性。
DOI：
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发表时间：
1980
影响因子：
4.8
通讯作者：
S. Springhorn
中科院分区：
文献类型：
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作者：
S. Lacks;S. Springhorn
许多酶，包括淀粉酶、淀粉酶和蛋白酶，在十二烷基硫酸钠存在下经聚丙烯酰胺凝胶电泳后显示可复性。通过对引入凝胶中的底物的作用以及随后对产物或未反应底物的染色来原位检测酶活性。该技术结合了酶鉴定的优势和十二烷基硫酸钠存在下凝胶电泳的分辨率和分子量依赖性。一旦洗涤剂从凝胶中扩散出来，酶似乎就恢复了活性。大多数单体酶甚至在其二硫键被破坏后也可以复性，但包括胰蛋白酶在内的几种蛋白酶却不能。由相同亚基组成的寡聚酶的复性性差。复性酶在电泳后保留在凝胶中的时间长于在没有洗涤剂的情况下进行电泳的天然酶。复性的酶的再电泳表明，保留的活性的一部分是物理锚定的凝胶，可能是通过折叠的多肽arrhenous凝胶基质作为酶的复性。
A number of enzymes, including amylases, dehydrogenases, and proteases, were shown to be renaturable after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Enzyme activity was detected in situ by action on substrates introduced into the gel and subsequent staining of either the product or unreacted substrate. This technique combines the advantage of enzyme identification with the resolution and molecular weight dependence of gel electrophoresis in the presence of sodium dodecyl sulfate. Enzymes appeared to recover activity as soon as the detergent diffused out of the gel. Most monomeric enzymes could be renatured even after disruption of their disulfide bonds, but several proteases, including trypsin, could not. Oligomeric enzymes composed of identical subunits were poorly renaturable. Renatured enzymes were retained in gels after electrophoresis longer than native enzymes which had been subjected to electrophoresis in the absence of detergent. Re-electrophoresis of the renatured enzymes showed that part of the retained activity was physically anchored to the gel, possibly by the folding of polypeptide aroung the gel matrix as the enzymes were renatured.