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pH difference across the outer mitochondrial membrane measured with a green fluorescent protein mutant.

Abstract：

:In this study we have generated a EYFP targeted to the mitochondrial intermembrane space (MIMS-EYFP) to determine for the first time the pH within this compartment. The fragment encoding HAI-tagged EYFP was fused with the C-terminus of glycerol-phosphate dehydrogenase, an integral protein of the inner mitochondrial membrane. Human ECV304 cells transiently transfected with MIMS-EYFP showed the typical mitochondrial network, co-localized with MitoTracker Red. Following the calibration procedure, an estimation of the pH value in the intermembrane space was obtained. This value (6.88+/-0.09) was significantly lower than that determined in the cytosol after transfection with a cytosolic EYFP (7.59+/-0.01). Further, the pH of the mitochondrial matrix, determined with a EYFP targeted to this subcompartment, was 0.9 pH units higher than that in the intermembrane space. In conclusion, MIMS-EYFP represents a novel powerful tool to monitor pH changes in the mitochondrial intermembrane space of live cells.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Porcelli AM,Ghelli A,Zanna C,Pinton P,Rizzuto R,Rugolo M

doi

10.1016/j.bbrc.2004.11.105

subject

Has Abstract

pub_date

2005-01-28 00:00:00

pages

799-804

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(04)02689-0

journal_volume

326

pub_type

杂志文章

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