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Titolo:
RECONSTITUTION OF THE F-0 COMPLEX OF ESCHERICHIA-COLI ATP SYNTHASE FROM ISOLATED SUBUNITS - VARYING THE NUMBER OF ESSENTIAL CARBOXYLATES BYCO-INCORPORATION OF WILD-TYPE AND MUTANT SUBUNIT-C AFTER PURIFICATIONIN ORGANIC-SOLVENT
Autore:
DMITRIEV OY; ALTENDORF K; FILLINGAME RH;
Indirizzi:
UNIV WISCONSIN,SCH MED,DEPT BIOMOLEC CHEM,1300 UNIV AVE MADISON WI 53706 UNIV WISCONSIN,SCH MED,DEPT BIOMOLEC CHEM MADISON WI 53706 UNIV OSNABRUCK,FACHBEREICH BIOL CHEM W-4500 OSNABRUCK GERMANY
Titolo Testata:
European journal of biochemistry
fascicolo: 2, volume: 233, anno: 1995,
pagine: 478 - 483
SICI:
0014-2956(1995)233:2<478:ROTFCO>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
REACTIVE PROTEIN COMPONENT; ENERGY-TRANSDUCING SYSTEM; H+-ATPASE; UNC OPERON; F1F0; F0; DICYCLOHEXYLCARBODIIMIDE; TRANSLOCATION; MUTATIONS;
Keywords:
ATP SYNTHASE; F-0; PROTON TRANSLOCATION; SUBUNIT C;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
25
Recensione:
Indirizzi per estratti:
Citazione:
O.Y. Dmitriev et al., "RECONSTITUTION OF THE F-0 COMPLEX OF ESCHERICHIA-COLI ATP SYNTHASE FROM ISOLATED SUBUNITS - VARYING THE NUMBER OF ESSENTIAL CARBOXYLATES BYCO-INCORPORATION OF WILD-TYPE AND MUTANT SUBUNIT-C AFTER PURIFICATIONIN ORGANIC-SOLVENT", European journal of biochemistry, 233(2), 1995, pp. 478-483

Abstract

Subunit c of the Escherichia coli F1F0-ATPase, purified in chloroform/methanol (2:1), was reconstituted with detergent-solubilized F-0 subunits a and b to form a functionally active H+ channel. The rates of Huptake by the proteoliposomes containing the reconstituted F-0 complex were comparable to those observed with native F-0 reconstituted without subunit dissociation. The F-0 reconstituted from purified subunitswas also shown to form an active ATP-driven H+ pump upon binding of the F-1-ATPase sector of the complex. Reconstitution of D61N and D61G mutant c subunits with wild-type subunits a and b produced an inactive F-0. Hybrid F-0 complexes, formed with mixtures of wild-type and D61N or D61G mutant c subunits, were also prepared. Formation of an active F-0 was prevented by addition of relatively small proportions of D61N or D61G mutant c subunits, i.e. active F-0 formation was gradually disrupted as the mutant/wild-type ratio was increased from 0.05 to 0.2. The hybrid reconstitution studies support a model where inactivation ofone of the 9-12 c subunits found in F-0 is sufficient to abolish activity.

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Documento generato il 21/09/20 alle ore 01:24:10