The behavior of horse liver alcohol dehydrogenase in guanidine hydrochloride solutions.

نویسندگان

  • R W Green
  • R H McKay
چکیده

The z-average molecular weight of horse liver alcohol dehydrogenase in dilute neutral buffer is 78,200 by sedimentation equilibrium. Guanidine hydrochloride, 1 M, reversibly inhibits enzymatic activity. The inhibition with nicotinamide adenine dinucleotide is competitive and with ethanol is mixed. When ethylenediaminetetraacetate is added to the enzyme in 1 M guanidine hydrochloride, zinc is removed, followed by aggregation. Sedimentation velocity experiments indicate that no subunit is formed under these conditions. In 3 M guanidine hydrochloride, the sedimentation equilibrium data suggest the presence of a reversible associationdissociation system. Ethylenediaminetetraacetate removes zinc but has no effect on sedimentation equilibrium behavior of the enzyme in 3 M guanidine hydrochloride. Reduction and alkylation of the enzyme in 3 M guanidine hydrochloride results in almost complete dissociation into two subunits. Thus, it appears that sulfhydryl residues may be involved in subunit association, whereas zinc plays little or no role. After correction for guanidine binding, the z-average molecular weight of the enzyme in 5 M guanidine hydrochloride containing mercaptoethanol is 41,000. Measurements of molecular weight, sedimentation coefficient, and intrinsic viscosity of the enzyme in 5 M guanidine hydrochloride all indicate that the structure of horse liver alcohol dehydrogenase is composed of two subunits.

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عنوان ژورنال:
  • The Journal of biological chemistry

دوره 244 18  شماره 

صفحات  -

تاریخ انتشار 1969