Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form.

Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Research Abstract Details 

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Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Abstract Text:

N Ohtani,M Haruki,A Muroya,M Morikawa,S Kanaya,

Escherichia coli RNase HII is composed of 198 amino acid residues. The enzyme has been overproduced in an insoluble form, purified in a urea-denatured form, and refolded with poor yield [M. Itaya (1990) Proc. Natl. Acad. Sci. USA 87, 8587-8591]. To facilitate the preparation of the enzyme in an amount sufficient for physicochemical studies, we constructed an overproducing strain in which E. coli RNase HII is produced in a soluble form. The enzyme was purified from this strain and its biochemical and physicochemical properties were characterized. The good agreement in the molecular weights estimated from SDS-PAGE (23,000) and gel filtration (22,000) suggests that the enzyme acts as a monomer. From the far-UV circular dichroism spectrum, its helical content was calculated to be 23%. The enzyme showed Mn(2+)-dependent RNase H activity. Its specific activity determined using (3)H-labeled M13 RNA/DNA hybrid as a substrate was comparable to but slightly higher than that of the refolded enzyme, indicating that the enzyme overproduced and purified in a soluble form is more suitable for structural and functional analyses than the refolded enzyme.

Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Publishing Authors By Initials

N Ohtani,M Haruki,A Muroya,M Morikawa,S Kanaya,

For similar natural sciences: chemistry: chemistry, physical: solubility research abstracts see: natural sciences: chemistry: chemistry, physical: solubility research

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Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 127

Page Numbers: 895-9

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: May

YEAR: 2000

Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Keywords Mesh Terms:

KEYWORDS: Solubility

MESH TERMS: metabolism

Chemical & Substance for Abstract: Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. Information

Substance Name: ribonuclease HI

Registry Number: EC 3.1.26.4

Grant and Affiliation Information for Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form.

AFFILIATION: Department of Material and Life Science, Graduate School of Engineering, Osaka University, Yamadaoka, Suita, Osaka 565-0871, Japan.

Country: JAPAN

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MEDLINETA: J Biochem

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