Active Subunits of Transaldolase Bound to Sepharose

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The properties of subunits of avidin coupled to sepharose.

Avidin that had been coupled to Sepharose 4B activated with CNBr retained over 90% of its biotin-binding capacity. When low concentrations of CNBr were used about 75% of the protein could be removed from the Sepharose by washing with guanidinium chloride (6 m). The remaining 25%, the covalently bound subunits, had an almost undiminished capacity for biotin but a decreased affinity. Addition of ...

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Isolation of rabbit reticulocyte initiation factors by means of heparin bound to sepharose.

Passage of cell-free extracts of rabbit reticulocytes through heparin-Sepharose affinity columns results in the loss of the ability of the effluent to initiate protein synthesis. This is shown by the loss of response to added rabbit globin mRNA or to inhibitors of initiation of protein synthesis, such as heparin and aurin tricarboxylic acid, and by recovery of initiation activity by addition of...

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Purification of the subunits of transcarboxylase by affinity chromatography on avidin-sepharose.

Transcarboxylase consists of a central 12 SH subunits each of which is linked to the central subunit by two similar to 1.3 SE biotin carboxyl carrier proteins. The subunits from dissociated transcarboxylase have been difficult to isolate because conditions which stabilize them also promote their reassociation to the intact enzyme. In this paper, we describe the use of avidin-Sepharose to adsorb...

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Separation of ribosomal subunits of Escherichia coli by Sepharose chromatography using reverse salt gradient.

A mixture of 30 S and 50 S subunits quantitatively absorbs on a column of Sepharose--4B from the buffer: 0.02 M Tris--HCl, pH 7.5, containing 1.5 M (NH4)2SO4. During elution by reverse gradient of ammonium sulphate (1.5--0.05 M) the subunits are eluted at different salt concentrations. Complete separation of subunits is attained in the absence of Mg2+ ions. The 30 S subunits prepared from 70 S ...

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All factors required for protein synthesis are retained on heparin bound to Sepharose.

1. Postmitochondrial supernatants of rabbit reticulocyte lysates were chromatographed on heparin bound to Sepharose 4B, and the fraction retained on affinity columns was separated by subsequent gel filtration on Sepharose 4B into three fractions, two of them active in protein synthesis. 2. The heavier fraction sedimented at 40S and contained more than 10% RNA. This consisted predominantly of a ...

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ژورنال

عنوان ژورنال: European Journal of Biochemistry

سال: 1973

ISSN: 0014-2956,1432-1033

DOI: 10.1111/j.1432-1033.1973.tb03224.x