Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form.

Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Research Abstract Details 

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Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Abstract Text:

K Sato,Y Nishina,K Shiga,

Electron-transferring flavoprotein (ETF) isolated from pig kidney is a heterodimer containing one FAD and one AMP [Sato, K. et al. (1993) J. Biochem. 114, 215-222]. This paper presents a method for separation of the alpha and beta subunits on a preparative scale. The subunits were separated by cation-exchange chromatography in the presence of 6 M urea at pH 7.6. The subunits were obtained in unfolded forms with denaturant. The unfolded subunits were restored to the heterodimeric form having FAD and AMP with high yield after incubation with FAD and AMP under non-denaturing conditions. The far-UV CD spectrum, the fluorescence and absorption spectra of the bound FAD, and the enzyme activity of the reconstituted ETF were all identical to those of the ETF isolated from pig kidney. Thus, an experimental system was established for the reconstitution of ETF from the four components, alpha, beta, FAD, and AMP. Analyses of the absorption spectra of the separated subunits suggested that the contents of aromatic residues were one tryptophan and six tyrosines in alpha and one tryptophan and one tyrosine in beta. The possibility that one subunit binds with FAD or AMP in the absence of the other subunit was examined by measuring the flavin and protein fluorescence spectra, but no spectral changes reflecting such binding were detected. This result suggests that the coexistence of both the subunits is necessary for the binding of FAD and AMP with the protein.

Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Publishing Authors By Initials

K Sato,Y Nishina,K Shiga,

For similar animals: chordata: vertebrates: mammals: artiodactyla: swine research abstracts see: animals: chordata: vertebrates: mammals: artiodactyla: swine research

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Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 116

Page Numbers: 147-55

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jul

YEAR: 1994

Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Keywords Mesh Terms:

KEYWORDS: Swine

MESH TERMS: isolation & purification

Chemical & Substance for Abstract: Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form. Information

Substance Name: Adenosine Monophosphate

Registry Number: 61-19-8

Grant and Affiliation Information for Preparation of separated alpha and beta subunits of electron-transferring flavoprotein in unfolded forms and their restoration to the native holoprotein form.

AFFILIATION: Department of Physiology, Kumamoto University School of Medicine.

Country: JAPAN

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MEDLINETA: J Biochem

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