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Thiophosphate as a probe for mitochondrial oxidative phosphorylation, model phosphorylation reactions and membrane permeability

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Abstract

  1. 1.

    Monothiophosphate was used as a probe for substrate level phosphorylation, mitochondrial oxidative phosphorylation, and membrane permeability.

  2. 2.

    Thiophosphate does not support ATP synthesis either in mitochondria or model enzymic reactions, conservation of oxidative energy occurring through formation of ATP(γ)S.

  3. 3.

    Thiophosphoryl transfer is slower compared to phosphoryl (group) transfer. Such behavior is compatible with an addition-elimination mechanism.

  4. 4.

    Thiophosphate accumulates in mitochondria by an energydependent process and substitutes for phosphate in the catalysis of metabolite transport. Relative to phosphate, a slower and less extensive permeation was observed.

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Abbreviations

TMPD:

N,N,N',N'-tetramethyl-p-phenylene-diamine dihydrochloride

MES:

2 (N-morpholino)ethane sulfonic acid

TES:

N-tris (Hydroxymethyl) methyl-2-aminomethane sulfonic acid

PEI:

polyethyleneimine

ATP(γ)S:

thiophosphoryl ADP

DTT:

dithiothreitol

RLM:

rat liver mitochondria

SP:

thiophosphate

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

PGK:

phosphoglycerate kinase

MOPS:

morpholinopropane sulfonic acid.

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Author notes

  1. Gregory R. Schonbaum

    Present address: Department of Biochemistry, St. Jude's Children's Research Hospital, 38101, Memphis, Tennessee

Authors and Affiliations

  1. Department of Physiology-Anatomy, University of California, 94720, Berkeley, California

    Joaquim Tavares de Sousa, Lester Packer & Gregory R. Schonbaum

Authors
  1. Joaquim Tavares de Sousa
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  2. Lester Packer
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  3. Gregory R. Schonbaum
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Tavares de Sousa, J., Packer, L. & Schonbaum, G.R. Thiophosphate as a probe for mitochondrial oxidative phosphorylation, model phosphorylation reactions and membrane permeability. J Bioenerg Biomembr 3, 539–552 (1972). https://doi.org/10.1007/BF01539063

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  • DOI: https://doi.org/10.1007/BF01539063

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