Abstract

Nephropathic cystinosis is an autosomal recessive disease characterized by intra-lysosomal accumulation of the disulfide amino acid cystine. Our previous work has shown that skin fibroblasts cultured from patients with cystinosis accumulate lysosomal cystine from the degration of both exogenous and endogenous disulfide containing proteins. We propose to employ these fibroblasts to study the disulfide reduction and degradation of endogenous and exogenous proteins to determine the relative amount of lysosomal participation in the degradation of each class of proteins. These studies will require production of 35(S)-labelled rat serum albumin, and 35(S)-insulin. The results of the interaction of these uniquely labelled proteins with cystinotic fibroblasts will be analyzed both by SDS polyacrylamide gel electrophoresis, high performance liquid chromatography using coupled UV and radio-chemical detectors, and GC-mass spectrometry. This project should provide data relevant to several fundamental questions regarding cellular proteolysis from a novel perspective.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK025548-06
Application #
3227481
Study Section
Biochemistry Study Section (BIO)
Project Start
1979-12-01
Project End
1986-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109

  Publications

Shotelersuk, V; Larson, D; Anikster, Y et al. (1998) CTNS mutations in an American-based population of cystinosis patients. Am J Hum Genet 63:1352-62
Pisoni, R L; Park, G Y; Velilla, V Q et al. (1995) Detection and characterization of a transport system mediating cysteamine entry into human fibroblast lysosomes. Specificity for aminoethylthiol and aminoethylsulfide derivatives. J Biol Chem 270:1179-84
Pisoni, R L; Lemons, R M; Paelicke, K M et al. (1992) Description of a selection method highly cytotoxic for cystinotic fibroblasts but not normal human fibroblasts. Somat Cell Mol Genet 18:1-6
de Cespedes, C; Thoene, J G; Lowler, K et al. (1992) Leucine and tissue distribution of bulky and small neutral amino acids in rats: dissociation between transport and insulin-mediated effects. J Inherit Metab Dis 15:145-54
Lemons, R M; Thoene, J G (1991) Mediated calcium transport by isolated human fibroblast lysosomes. J Biol Chem 266:14378-82
Pisoni, R L; Thoene, J G (1991) The transport systems of mammalian lysosomes. Biochim Biophys Acta 1071:351-73
Pisoni, R L; Acker, T L; Lisowski, K M et al. (1990) A cysteine-specific lysosomal transport system provides a major route for the delivery of thiol to human fibroblast lysosomes: possible role in supporting lysosomal proteolysis. J Cell Biol 110:327-35
Pisoni, R L; Lisowski, K M; Lemons, R M et al. (1989) Utilization of mercaptoethylgluconamide for depleting human cystinotic fibroblasts of their accumulated lysosomal cystine. Pediatr Res 26:73-6
Pisoni, R L; Thoene, J G (1989) Detection and characterization of a nucleoside transport system in human fibroblast lysosomes. J Biol Chem 264:4850-6
de Cespedes, C; Thoene, J G; Lowler, K et al. (1989) Evidence for inhibition of exodus of small neutral amino acids from non-brain tissues in hyperphenylalaninaemic rats. J Inherit Metab Dis 12:166-80

Showing the most recent 10 out of 13 publications