The Effect of Triton WR-1339 on the Subcellular Distribution of Trypan Blue and 125 I-labelled Albumin in Rat Liver

Overview

Journal Biochem J

Specialty Biochemistry

Date 1973 Sep 1

PMID 4772628

Authors

M Davies

Affiliations

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Abstract

1. The density-gradient distribution patterns of acid phosphatase, Trypan Blue and denatured (125)I-labelled albumin were studied by discontinuous sucrose- and isopycnic sucrose-density-gradient centrifugation on combined heavy and light mitochondrial (M+L) fractions of liver isolated from normal rats and from rats injected with Triton WR-1339. 2. The results obtained from the subfractionation of the M+L pellet of normal animals indicate that the equilibrium density of Trypan Blue and acid-insoluble radioactivity is the same as that for acid phosphatase, which suggests they are bound by a common membrane to form a distinct subcellular population of lysosomal nature. 3. In contrast, the analysis of the isopycnic gradients obtained on subfractionation of M+L pellets of liver isolated from rats treated with Triton WR-1339 show that the acid-insoluble radioactivity has an equilibrium density around 1.21, whereas the acid hydrolases, including cathepsin D, show the characteristic shift to an equilibrium density of around 1.12. Trypan Blue is distributed along the gradient with distinct peaks at densities 1.22 and 1.12. 4. Similar equilibrium-density distribution patterns were obtained with M+L pellets isolated from rats pretreated with Triton WR-1339 but not injected with Trypan Blue. 5. Treatment of the rats with Triton WR-1339 does not affect albumin digestion of isolated intact lysosomes despite the fact that most of the cathepsin D and the albumin ingested by phagocytosis are located in different vacuoles. 6. It is concluded from these experiments that in the liver of animals treated with Triton WR-1339 (125)I-labelled albumin is located within heterophagosomes which do not fuse with heterolysosomes containing the non-ionic detergent Triton WR-1339. The inability of these two lysosomal populations to fuse is not due to Trypan Blue.

References

Norton W, Lewis D, ZIFF M . Electron-dense deposits following injection of gold sodium thiomalate and thiomalic acid. Arthritis Rheum. 1968; 11(3):436-43. DOI: 10.1002/art.1780110309. View

NEIL M, Horner M . Studies on acid hydrolases in adult and foetal tissues. Acid rho-nitrophenyl phosphate phosphohydrolases of adult guinea-pig liver. Biochem J. 1964; 92(1):217-24. PMC: 1215459. DOI: 10.1042/bj0920217. View

Ennis R, Granda J, Posner A . Effect of gold salts and other drugs on the release and activity of lysosomal hydrolases. Arthritis Rheum. 1968; 11(6):756-64. DOI: 10.1002/art.1780110605. View

Gregoriadis G, Morell A, Sternlieb I, Scheinberg I . Catabolism of desialylated ceruloplasmin in the liver. J Biol Chem. 1970; 245(21):5833-7. View

Leighton F, Poole B, BEAUFAY H, Baudhuin P, Coffey J, Fowler S . The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968; 37(2):482-513. PMC: 2107417. DOI: 10.1083/jcb.37.2.482. View

BEAUFAY H, Jacques P, Baudhuin P, Sellinger O, Berthet J, DE DUVE C . Tissue fractionation studies. 18. Resolution of mitochondrial fractions from rat liver into three distinct populations of cytoplasmic particles by means of density equilibration in various gradients. Biochem J. 1964; 92(1):184-205. PMC: 1215457. DOI: 10.1042/bj0920184. View

Davies M, Lloyd J, Beck F . The effect of Trypan Blue, suramin and aurothiomalate on the breakdown of 125 I-labelled albumin within rat liver lysosomes. Biochem J. 1971; 121(1):21-6. PMC: 1176481. DOI: 10.1042/bj1210021. View

Gregoriadis G, RYMAN B . Lysosomal localization of -fructofuranosidase-containing liposomes injected into rats. Biochem J. 1972; 129(1):123-33. PMC: 1174048. DOI: 10.1042/bj1290123. View

Lloyd J, Beck F . The relationship of chemical structure to teratogenic activity among bisazo dyes: a re-evaluation. J Embryol Exp Morphol. 1966; 16(1):29-39. View

10.

WILLS E . Enzyme inhibition by suramin and the measurement of the isoelectric points of some enzymes. Biochem J. 1952; 50(3):421-5. PMC: 1197671. DOI: 10.1042/bj0500421. View

11.

TORRIANI A . Influence of inorganic phosphate in the formation of phosphatases by Escherichia coli. Biochim Biophys Acta. 1960; 38:460-9. DOI: 10.1016/0006-3002(60)91281-6. View

12.

Mego J, McQUEEN J . THE UPTAKE AND DEGRADATION OF INJECTED LABELED PROTEINS BY MOUSE-LIVER PARTICLES. Biochim Biophys Acta. 1965; 100:136-43. DOI: 10.1016/0304-4165(65)90436-8. View

13.

Mego J, Bertini F, McQUEEN J . The use of formaldehyde-treated 131-I-albumin in the study of digestive vacuoles and some properties of these particles from mouse liver. J Cell Biol. 1967; 32(3):699-707. PMC: 2107274. DOI: 10.1083/jcb.32.3.699. View

14.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

15.

DINGLE J . Vacuoles, vesicles and lysosomes. Br Med Bull. 1968; 24(2):141-5. DOI: 10.1093/oxfordjournals.bmb.a070616. View

16.

Persellin R, ZIFF M . The effect of gold salt on lysosomal enzymes of the peritoneal macrophage. Arthritis Rheum. 1966; 9(1):57-65. DOI: 10.1002/art.1780090107. View