SYSTEMATIC STUDY OF EFFECTS OF NONIONIC DETERGENTS ON SOLUBILIZATION AND ACTIVITY OF PIG KIDNEY ADENYLATE-CYCLASE
SYSTEMATIC STUDY OF EFFECTS OF NONIONIC DETERGENTS ON SOLUBILIZATION AND ACTIVITY OF PIG KIDNEY ADENYLATE-CYCLASE
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DOI：
10.1111/j.1432-1033.1978.tb12753.x
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发表时间：
1978-01-01
期刊：
影响因子：
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通讯作者：
JARD, S
中科院分区：
文献类型：
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作者：
GUILLON, G;ROY, C;JARD, S
研究了17种非离子洗涤剂对猪肾质膜腺苷酸环化酶[EC 4.6.1.1]的影响（洋地黄苷；Triton系列洗涤剂：X45、X100、X102、X114、X165、X305、X405、N101； Brij系列洗涤剂：35、56、96和Tween系列洗涤剂：20、40、60、80、85）。膜（2.66 mg蛋白/ml）在0℃下用越来越多的洗涤剂处理150 min。C.在30度下测量处理膜及其相应的不可沉淀部分中腺苷酸环化酶的活性。C.只有亲水亲脂平衡值在12-14之间的洗涤剂才是有效的增溶剂。对于这些洗涤剂，增溶率取决于它们的结构。溶解时腺苷酸环化酶保持在0度。C在30度孵育。C时，酶的催化活性呈指数下降，半衰期为5 min，达到平衡值。这种酶活性的温度依赖性调节是可逆的。其大小取决于所使用的洗涤剂。非增溶性洗涤剂的效果更明显，但不太明显。在培养培养基中，洗涤剂的胶束形式是导致溶酶分子群体中因底物可及性不同而出现微观异质性的原因。当降低培养介质中的洗涤剂浓度时，这种影响被抑制。
The effects of 17 non-ionic detergents on pig kidney plasma membrane adenylate cyclase [EC 4.6.1.1] were tested (digitonin; detergents from the Triton series: X45, X100, X102, X114, X165, X305, X405, N101; the Brij series: 35, 56, 96 and the Tween series: 20, 40, 60, 80, 85). Membranes (2.66 mg protein/ml) were treated with increasing amounts of detergent for 150 min at 0.degree. C. The adenylate cyclase activities present in treated membranes and in their corresponding non-sedimentable fractions were measured at 30.degree. C. Only detergents having hydrophilic lipophilic balance values from 12-14 were effective solubilizing agents. For these detergents, the solubilization yield was dependent on their structure. When solubilized adenylate cyclase kept at 0.degree. C was incubated at 30.degree. C, the enzyme catalytic activity exponentially decreased towards an equilibrium value with a half-time of 5 min. This temperature-dependent adjustment of enzyme activity was reversible. Its magnitude was dependent on the detergent used. A more progressive but less pronounced effect was observed with non-solubilizing detergents. The micellar form of detergent in the incubation medium was responsible for the appearance of microheterogeneity in the population of solubilized enzyme molecules differing by the substrate accessibility. This effect was suppressed when reducing the detergent concentration in the incubation medium.