Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases.

来自 dx.doi.org

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130

作者：

LM Siegel，KJ Monty

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摘要：

The behavior of each of a series of proteins during chromatography on columns of Sephadex G-200 may be correlated with the Stokes radius of the protein, but does not correlate with molecular weight. Proteins with Stokes radii as high as 107 or molecular weights as high as 1 300 000 may be characterized by the use of such columns. The Sephadex data are used in a critique of earlier mathematical treatments of the phenomenon known as "gel filtration". With a Stokes radius measured by the chromatographic method and a sedimentation coefficient determined by density gradient centrifugation, reasonable estimates for both the molecular weight and the frictional ratio ( f/ f 0) of a macromolecule are available. Since both of these methods are applicable to proteins present in mixtures, valuable information concerning the molecular weights and shapes of proteins may be obtained in anticipation of the achievement of high degrees of purity. The determination of the molecular weight and the f/ f 0 for each of several enzymes in unfractionated extracts of Salmonella typhimurium and Neurospora crassa illustrates this application.

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关键词：

Salmonella typhimurium Sulfites Cytochromes Oxidoreductases Alcohol Oxidoreductases Urease Proteins Serum Albumin Fibrinogen Centrifugation