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Slater 1953 Biochem J

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Slater EC, Cleland KW (1953) The effect of tonicity of the medium on the respiratory and phosphorylative activity of heart-muscle sarcosomes. Biochem J 53:557-67.

» PMID: 13032109 Open Access

Slater EC, Cleland KW (1953) Biochem J

Abstract:

  1. The α-ketoglutaric oxidase system of heartmuscle sarcosomes has a pH optimum at 7.4. The yield of oxidative phosphorylation (P:O ratio) is unchanged between pH 6.2 and 7.7.
  2. Hypertonic sucrose (0.88 M) is an inhibitor of the succinic oxidase system in the Keilin & Hartree preparation. Its major effect appears to be on the accessibility of both the endogenous and added cytochrome c to the other components of the system.
  3. Maximum activity of the α-ketoglutaric oxidase system of heart-muscle sarcosomes is obtained under the most highly hypotonic conditions studied, equivalent to about one-third isotonic. Under these conditions, sarcosomes are swollen, but shrink again when placed in isotonic medium. The effect of tonicity on the activity of the α-ketoglutaric oxidase system is also reversible.
  4. As the tonicity is increased by saline, sucrose or phosphate, the activity of the α-ketoglutaric oxidase system decreases.
  5. The P:O ratio is not affected over a wide range of sucrose concentrations which have a marked effect on the activity of the α-ketoglutaric oxidase system. This and other examples where the oxidase system is more sensitive than the P:O ratio to variations of the conditions indicates that the phosphorylative enzymes are normally in excess of the purely oxidative enzymes and increases the likelihood that measurements of the yield of oxidative phosphorylation on isolated tissue preparations represent the state of affairs in the cell.
  6. Phosphate, in high concentration, decreases the P:O ratio; the optimal concentration is 0.03 M.
  7. Hypertonic sucrose is unsuitable for the isolation of sarcosomes. There is, however, no significant difference between sarcosomes isolated with isotonic sucrose and isotonic saline, except that the latter are deficient in cytochrome c.

Keywords: Tonicity, α-ketoglutamate dehydrogenase


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Organism: Rat  Tissue;cell: Heart  Preparation: Isolated mitochondria 




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