昆虫线粒体外甘油磷酸脱氢酶II。蜜蜂胸脯酶的性质和氨基酸组成

Ronald W. Brosemer, Ronald R. Marquardt
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引用次数: 37

摘要

测定了结晶蜜蜂胸廓甘油磷酸脱氢酶(l-甘油-3-磷酸:DPN+氧化还原酶,EC 1.1.1.8)的几种酶学性质。磷酸二羟丙酮的表观米歇里斯常数为0.33 mM;与底物有关时,活性没有增加。蜜蜂酶在pH 6.6左右有广泛的最适pH值,而兔肌酶在pH 7.7左右有明显的最适pH值。蜜蜂酶在21°温度下从pH 4.8到9.9稳定15分钟。蜜蜂酶的温度系数Q10在21°36°范围内为1.7。酶在55°时稳定5min,在61°时完全失活。蜜蜂酶对2个DPN+类似物的相对反应性与兔酶相同。低浓度的对汞苯甲酸酯(PCMB)对蜜蜂酶有抑制作用,对n -乙基马来酰亚胺不太敏感,1 mM碘乙酸不受抑制。谷胱甘肽不能激活这种酶。蜜蜂酶的氨基酸组成与先前报道的兔肌酶的组成有很大不同。以1个色氨酸残基为基础的蜜蜂酶的最小分子量为32 700。由于在Sephadex G-200色谱柱上测定的分子量约为67 000,氨基酸组成表明摩尔重量为65 400。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Insect extramitochondrial glycerophosphate dehydrogenase II. Enzymic properties and amino acid composition of the enzyme from honeybee (Apis mellifera) thoraces

Several enzymic properties of crystalline honeybee (Apis mellifera) thoracic glycerophosphate dehydrogenase (L-glycerol-3-phosphate:DPN+ oxidoreductase, EC 1.1.1.8) were determined. The apparent Michaelis constant for dihydroxyacetone phosphate is 0.33 mM; there is no increase in activity with substrate concn. above 0.5 mM.

The bee enzyme has a broad pH optimum around pH 6.6, while the rabbits-muscle enzyme has a sharp optimum at pH 7.7. The bee enzyme is stable for 15 min at 21° from pH 4.8 to 9.9.

The temperature coefficient, Q10, for the bee enzyme is 1.7 in the range from 21° 36°. The enzyme is stable for 5 min at 55° and completely inactivated at 61°. The bee enzyme shows the same relative reactivity with 2 DPN+ analogues as does the rabbit enzyme. The bee enzyme is inhibited by a low concentration of p-mercuribenzoate (PCMB), is less sensitive to N-ethylmaleimide, and is not inhibited by 1 mM iodoacetate. Glutathione does not activate the enzyme.

The amino acid composition of the bee enzyme is quite different from the previously reported composition of the rabbits-muscle enzyme. The minimum molecular weight of the bee enzyme based on 1 tryptophan residue is 32 700. Since the molecular weight determined on a Sephadex G-200 column is around 67 000, the amino acid composition indicates a mol. wt. of 65 400.

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