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Effects of Tl+ on the inner membrane thiol groups, respiration, and swelling in succinate-energized rat liver mitochondria were modified by thiol reagents

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Abstract

The effects of both Tl+ and thiol reagents were studied on the content of the inner membrane free SH-groups, detected with Ellman reagent, and the inner membrane potential as well as swelling and respiration of succinate-energized rat liver mitochondria in medium containing TlNO3 and KNO3. These effects resulted in a rise in swelling and a decrease in the content, the potential, and mitochondrial respiration in 3 and 2,4-dinitrophenol-uncoupled states. A maximal effect was seen when phenylarsine oxide reacting with thiol groups recessed into the hydrophobic regions of the membrane. Compared with phenylarsine oxide, the effective concentrations of other reagents were approximately one order of magnitude higher in experiments with mersalyl and 4,4′-diisothiocyanostilbene-2,2′-disulfonate, and two orders of magnitude higher in experiments with tert-butyl hydroperoxide and diamide. The above effects of Tl+ and the thiol reagents became even more pronounced with calcium overload of mitochondria. However, the effects were suppressed by inhibitors of the mitochondrial permeability transition pore (cyclosporine A, ADP, and n-ethylmaleimide). These findings suggest that opening of the pore induced by Tl+ in the inner membrane can be dependent on the conformation state of the adenine nucleotide translocase, which depends on the activity of its thiol groups.

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Abbreviations

ΔΨmito :

Inner mitochondrial membrane potential

ANT:

Adenine nucleotide translocase

CsA:

Cyclosporine A

CyP-D:

Cyclophilin D

Diam:

Diamide

DIDS:

4,4′-Diisothiocyanostilbene-2,2′-disulfonate

DNP:

2,4-Dinitrophenol

DTNB:

5,5-Dithio-bis-nitrobenzoic acid

DTT:

Dithiothreitol

EDTA:

Ethylenediaminetetraacetic acid

EGTA:

Ethylene glycol-bis(β-aminoethyl ether) N,N,N′,N′-tetraacetic acid

IMM:

Inner mitochondrial membrane

MPTP:

Mitochondrial permeability transition pore

MSL:

Mersalyl

NEM:

n-Ethylmaleimide

PAO:

Phenylarsine oxide

PiC:

Mitochondrial phosphate carrier

RLM:

Rat liver mitochondria

SDS:

Sodium dodecyl sulfate

tBHP:

Tert-butyl hydroperoxide

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Acknowledgements

The author is grateful to Ms Terttu Kaustia for correcting the English as well as Irina V. Brailovskaya, Victor V. Furaev, and Boris N. Kormilitsyn for help in isolating mitochondria and estimation of the protein thiol content and the oxygen consumption rates in rat liver mitochondrial suspensions. The research was carried out within the state assignment of FASO of Russia (theme No. AAAA-A18-118012290142-9). Safranin fluorescence was measured using of Research Resource Center equipment for the physiological, biochemical and molecular-biological studies (Sechenov Institute of Evolutionary Physiology and Biochemistry, the Russian Academy of Sciences).

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Correspondence to Sergey M. Korotkov.

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Korotkov, S.M. Effects of Tl+ on the inner membrane thiol groups, respiration, and swelling in succinate-energized rat liver mitochondria were modified by thiol reagents. Biometals 34, 987–1006 (2021). https://doi.org/10.1007/s10534-021-00329-6

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  • DOI: https://doi.org/10.1007/s10534-021-00329-6

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