[Effect Of Polyelectrolytes on Catalytic Activity of Alcohol Dehydrogenase].

A V Dubrovsky, E V Musina, A L Kim, S A Tikhonenko
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Abstract

Fluorescent and optical spectroscopy were used to study the interaction of alcohol dehydrogenase (ADH) with negatively charged polystyrene sulfonate (PSS) and dextran sulfate (DS), as well as positively charged poly(diallyldimethylammonium) (PDADMA). As found, DS and PDADMA did not affect the structural and catalytic enzyme properties. In contrast, PSS slightly decreased the protein self-fluorescence over 1 h of incubation, which is associated with partial destruction of its quaternary (globular) structure. Investigation of the ADH activity with and without PSS showed its dependency on the incubation time and the PSS presence. Sodium chloride (2.0 M and 0.2 M) or ammonium sulfate (0.1 M) added to the reaction mixture did not completely protect the enzyme quaternary structure from the PSS action. However ammonium sulfate or 0.2 M sodium chloride stabilized the enzyme and partially inhibited the negative PSS effect.

聚电解质对醇脱氢酶催化活性的影响
利用荧光光谱和光谱学研究了乙醇脱氢酶(ADH)与带负电荷的聚苯乙烯磺酸盐(PSS)和硫酸葡聚糖(DS)以及带正电荷的聚二烯基二甲基铵(PDADMA)的相互作用。结果发现,DS和PDADMA对结构和催化酶性质没有影响。相比之下,PSS在孵育1小时内略微降低了蛋白质的自荧光,这与其四级(球形)结构的部分破坏有关。对有PSS和不含PSS的ADH活性的研究表明,ADH活性与孵育时间和PSS的存在有关。在反应混合物中加入氯化钠(2.0 M和0.2 M)或硫酸铵(0.1 M)并不能完全保护酶的季型结构免受PSS的作用。硫酸铵和0.2 M氯化钠稳定了酶的活性,部分抑制了PSS的负作用。
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