Active Form ofPseudomonas mevalonii3-Hydroxy-3-methylglutaryl Coenzyme A Reductase

Kenneth S. Rogers , Victor W. Rodwell , Paul Geiger
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引用次数: 9

Abstract

Based on multiple gel permeation chromatographic experiments, we report a Stokes radius of 59.7 Å forPseudomonas mevalonii3-hydroxy-3methylglutaryl coenzyme A reductase (HMG-CoA reductase; EC 1.1.1.88) and its His381Asn, His381Gln, and His381Lys mutant enzymes. Comparison of this Stokes radius with the radius calculated from the crystal structure indicated that the active form ofP. mevaloniiHMG-CoA reductase was a hexamer and not a dimer as previously thought. The Stokes radius, anS20,wof 11.0, and an estimatedV̄of 0.723 were used in the Svedberg equation to calculate an anhydrous molecular mass of 270,084 Da forP. mevaloniiHMG-CoA reductase (monomer mass 45,538 Da), consistent with the enzyme being a hexamer in solution. The Stokes radii of all standard proteins examined correlated with the inverse error function complement of their partition coefficient,Kd.Kddid not correlate with logarithm of the standard protein's molecular weight. Eight nonstandard proteins had Stokes radii that matched their crystallographic radii of longest axis. This indicated that the frozen conformation of a protein in its crystal form can dictate restraints on its shape in solution.

甲瓦洛尼假单胞菌3-羟基-3-甲基戊二酰辅酶A还原酶的活性形式
基于多次凝胶渗透色谱实验,我们报道了mevalonii3-羟基-3甲基戊二酰辅酶a还原酶(HMG-CoA还原酶)的Stokes半径为59.7 Å;EC 1.1.1.88)及其His381Asn、His381Gln和His381Lys突变酶。将该Stokes半径与晶体结构计算的半径进行比较,表明p。mevaloniiHMG-CoA还原酶是六聚体,而不是以前认为的二聚体。在Svedberg方程中使用Stokes半径anS20,wof 11.0和估计的v ^ 0.723来计算p的无水分子质量为270,084 Da。mevaloniiHMG-CoA还原酶(单体质量45,538 Da),与该酶在溶液中为六聚体一致。所检测的所有标准蛋白的Stokes半径与其配分系数Kd的逆误差函数补相关。kd与标准蛋白分子量的对数无关。8种非标准蛋白质的斯托克斯半径与其最长轴的晶体学半径相匹配。这表明蛋白质在其晶体形式中的冷冻构象可以决定其在溶液中的形状限制。
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