Substitutional mutations in the uncoupling protein-specific sequences of mitochondrial uncoupling protein UCP1 lead to the reduction of fatty acid-induced H(+) uniport.

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F03%3A20030011" target="_blank" >RIV/67985823:_____/03:20030011 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Substitutional mutations in the uncoupling protein-specific sequences of mitochondrial uncoupling protein UCP1 lead to the reduction of fatty acid-induced H(+) uniport.

Popis výsledku v původním jazyce

Mutants were constructed for mitochondrial uncoupling protein UCP1, with single or multiple substitutions within or nearby the UCP-signatures located in the 1st alfa-helix and 2nd matrix segment, using the QuickChange site directed mutagenesis protocol (Stratagene), and were assayed fluorometrically for kinetics of fatty acid (FA)-induced H+ uniport and for Cl- uniport. Their ability to bind 3H-GTP was also evaluated. The wild type UCP1 was associated with the FA-induced H+ uniport proportional to the added protein with a Km for lauric acid of 43 mikroM and Vmax of 18 mikromol.min-1.(mg protein)-1. Neutralization of Arg 152 (in the 2nd matrix segment UCP-signature) led to ~50% reduction of FA affinity (reciprocal Km) and of Vmax for FA-induced H+ uniport. Halved FA-affinity and 70% reduction of Vmax was found for the double His substitution outside the signature (H145L & H147L mutant). Neutralization of Asp 27 in the1st alfa-helix UCP-signature (D27V mutant) resulted in 75% reduction o

Název v anglickém jazyce

Substitutional mutations in the uncoupling protein-specific sequences of mitochondrial uncoupling protein UCP1 lead to the reduction of fatty acid-induced H(+) uniport.

Popis výsledku anglicky

Mutants were constructed for mitochondrial uncoupling protein UCP1, with single or multiple substitutions within or nearby the UCP-signatures located in the 1st alfa-helix and 2nd matrix segment, using the QuickChange site directed mutagenesis protocol (Stratagene), and were assayed fluorometrically for kinetics of fatty acid (FA)-induced H+ uniport and for Cl- uniport. Their ability to bind 3H-GTP was also evaluated. The wild type UCP1 was associated with the FA-induced H+ uniport proportional to the added protein with a Km for lauric acid of 43 mikroM and Vmax of 18 mikromol.min-1.(mg protein)-1. Neutralization of Arg 152 (in the 2nd matrix segment UCP-signature) led to ~50% reduction of FA affinity (reciprocal Km) and of Vmax for FA-induced H+ uniport. Halved FA-affinity and 70% reduction of Vmax was found for the double His substitution outside the signature (H145L & H147L mutant). Neutralization of Asp 27 in the1st alfa-helix UCP-signature (D27V mutant) resulted in 75% reduction o

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2003

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

International Journal of Biochemistry & Cell Biology

ISSN

1357-2725

e-ISSN

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Svazek periodika

35

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

9

Strana od-do

212-220

Kód UT WoS článku

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EID výsledku v databázi Scopus

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