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PMID: 4853970 Published · ppublish English Journal Article

Purification and subunit analysis of wheat-germ ribonucleic acid polymerase II.

The Biochemical journal ·Vol. 139 ·No. 3 ·1974-06-00 ·Pages 771-7

Jendrisak JJ, Becker WM

Abstract

A procedure is described for the purification of the alpha-amanitin-sensitive DNA-dependent RNA polymerase [EC 2.7.7.6] from wheat germ. Solubilization of the enzyme activity was achieved by sonication of a crude extract in a high-salt buffer. Purification involved precipitation with protamine sulphate and (NH(4))(2)SO(4), chromatography on DEAE-cellulose and phosphocellulose, and sucrose gradient centrifugation. Under denaturing conditions the enzyme dissociated into five polypeptides with molecular weights and molar ratios of 220000 (0.9), 170000 (0.1), 140000 (1.0), 45000 (0.2), and 40000 (0.4). Approx. 1mg of purified RNA polymerase was obtained as a routine from 100g of starting material.

MeSH Terms
Adenosine Triphosphate Ammonium Sulfate Centrifugation, Density Gradient Chemical Precipitation Chromatography, DEAE-Cellulose DNA DNA-Directed RNA Polymerases/analysis,isolation & purification Electrophoresis, Polyacrylamide Gel Molecular Weight Nucleotides Protamines Protein Conformation Seeds/enzymology Triticum/enzymology Ultrasonics
Chemicals
Nucleotides Protamines Adenosine Triphosphate DNA DNA-Directed RNA Polymerases Ammonium Sulfate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Jendrisak J J
Becker W M
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20 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1390771
Published
1974-06-00
Pages
771-7
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1166341
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1390771 PMC Full Text View journal details
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