Purification and characterization of a thiol:protein disulfide oxidoreductase from bovine liver. 1977

D F Carmichael, and J E Morin, and J E Dixon

A thiol:protein disulfide oxidoreductase which degrades insulin into its A and B chains has been purified to homogeneity from bovine liver. The initial extraction procedure used 1% Triton X-100, which increased the soluble insulin degrading activity 100 to 800% as a result of solubilizing a membrane-associated enzyme. The criteria for homogeneity of the isolated protein include sodium dodecyl sulfate and disc gel electrophoresis. Characterization of the homogeneous protein by sedimentation equilibrium centrifugation indicates that a single protein is present which has a mass of 60,000 daltons. A single amino-terminal end group and molecular weight estimation by sodium dodecyl sulfate gel electrophoresis indicated that the thiol:protein disulfide oxidoreductase consists of a single polypeptide chain. The pure enzyme was also examined by gel filtration chromatography and has an apparent molecular weight of 92,000. The amino acid composition and carbohydrate content were also determined. The carbohydrate analysis demonstrated the presence of D-mannose, D-galactose, 2-acetamido-2-deoxy-D-glucose, and N-acetylneuraminic acid which comprise a total of 12% by weight of the isolated protein.

UI MeSH Term Description Entries
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D010088 Oxidoreductases The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9) Dehydrogenases,Oxidases,Oxidoreductase,Reductases,Dehydrogenase,Oxidase,Reductase
D011490 Protein Disulfide Reductase (Glutathione) An enzyme that catalyzes the reduction of a protein-disulfide in the presence of glutathione, forming a protein-dithiol. Insulin is one of its substrates. EC 1.8.4.2. Glutathione Insulin Transhydrogenase,Glutathione Protein Disulfide Oxidoreductase,Thiol-Disulfide Oxidoreductase,Thiol-Protein Disulfide Oxidoreductase,Disulfide Oxidoreductase, Thiol-Protein,Insulin Transhydrogenase, Glutathione,Oxidoreductase, Thiol-Disulfide,Oxidoreductase, Thiol-Protein Disulfide,Thiol Disulfide Oxidoreductase,Thiol Protein Disulfide Oxidoreductase,Transhydrogenase, Glutathione Insulin
D002417 Cattle Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor. Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D006601 Hexoses MONOSACCHARIDES whose molecules contain six carbon atoms, such as GLUCOSE and FRUCTOSE. They generally have the chemical formula C6H12O6. Hexose
D000596 Amino Acids Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins. Amino Acid,Acid, Amino,Acids, Amino
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012794 Sialic Acids A group of naturally occurring N-and O-acyl derivatives of the deoxyamino sugar neuraminic acid. They are ubiquitously distributed in many tissues. N-Acetylneuraminic Acids,Acids, N-Acetylneuraminic,Acids, Sialic,N Acetylneuraminic Acids

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