Heterogeneity of ADP diffusion and regulation of respiration in cardiac cells.

Abstract:

:Heterogeneity of ADP diffusion and regulation of respiration were studied in permeabilized cardiomyocytes and cardiac fibers in situ and in silico. Regular arrangement of mitochondria in cells was altered by short-time treatment with trypsin and visualized by confocal microscopy. Manipulation of matrix volumes by changing K(+) and sucrose concentrations did not affect the affinity for ADP either in isolated heart mitochondria or in skinned fibers. Pyruvate kinase (PK)-phosphoenolpyruvate (PEP) were used to trap ADP generated in Ca,MgATPase reactions. Inhibition of respiration by PK-PEP increased 2-3 times after disorganization of regular mitochondrial arrangement in cells. ADP produced locally in the mitochondrial creatine kinase reaction was not accessible to PK-PEP in intact permeabilized fibers, but some part of it was released from mitochondria after short proteolysis due to increased permeability of outer mitochondrial membrane. In in silico studies we show by mathematical modeling that these results can be explained by heterogeneity of ADP diffusion due to its restrictions at the outer mitochondrial membrane and in close areas, which is changed after proteolysis. Localized restrictions and heterogeneity of ADP diffusion demonstrate the importance of mitochondrial functional complexes with sarcoplasmic reticulum and myofibrillar structures and creatine kinase in regulation of oxidative phosphorylation.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Saks V,Kuznetsov A,Andrienko T,Usson Y,Appaix F,Guerrero K,Kaambre T,Sikk P,Lemba M,Vendelin M

doi

10.1016/S0006-3495(03)70065-4

subject

Has Abstract

pub_date

2003-05-01 00:00:00

pages

3436-56

issue

5

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(03)70065-4

journal_volume

84

pub_type

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