Structural Studies of Bovine Liver Rhodanese

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Improved preparation of bovine liver rhodanese.

Chemical studies of rhodanese (thiosulfate : cyanide sulfurtransferase, EC 2.8.1.1) in this laboratory have prompted the development of a modified procedure for preparing the crystalline enzyme. The new method, which is based on the earlier procedure of S6rbo (1) as modified by Westley et al. (2-4), permits isolation in apparently monodisperse form of nearly 50% of the total rhodanese activity ...

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Structural studies of bovine liver rhodanese. I. Isolation and characterization of two active forms of the enzyme.

Crystalline bovine liver rhodanese, prepared by ammonium sulfate and pH precipitation, has been shown to be comprised of two fully active components present in approximately equal amounts which are separable by polyacrylamide gel electrophoresis and by ion exchange chromatography. The two rhodanese forms, designated A and B on the basis of their order of elution from columns of DEAE-Sephadex, a...

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Dimeric structure and zinc content of bovine liver rhodanese.

Evidence from gel filtration and sedimentation studies of crystalline bovine liver rhodanese indicated that the enzyme molecule of 37,000 molecular weight is a dimer. In the native state, this form is in rapid, pa-dependent equilibrium with the monomeric species. A stable dimer is formed under mild oxidizing conditions. Analysis of the protein by peptide mapping indicated that the monomers are ...

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Acid pH-induced conformational changes in bovine liver rhodanese.

The enzyme rhodanese is greatly stabilized in the range pH 4-6, and samples at pH 5 are fully active after several days at 23 degrees C. This is very different from results at pH greater than 7, where there is significant loss of activity within 1 h. A pH-dependent conformational change occurs below pH 4 in a transition centered around pH 3.25 that leads slowly to inactive rhodanese at pH 3 (t ...

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Studies on the active site of rhodanese.

The complete loss of enzymic activity on reaction with alkylating agents, aromatic nitro compounds, or aliphatic mercaptans indicates the importance of sulfhydryl groups for catalysis. Analyses during the course of inactivation with any of these reagents revealed the loss of one of the two -SH groups in the rhodanese monomer when inactivation was complete. Amino acid analysis of iodoacetate-ina...

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ژورنال

عنوان ژورنال: Journal of Biological Chemistry

سال: 1971

ISSN: 0021-9258

DOI: 10.1016/s0021-9258(18)62306-3