The accurate delivery of newly synthesized and hydrolases from the rough endoplasmic reticulum to residency in lysosomes or extracellular fluids involves a series of co- and post-translational modifications of the protein and carbohydrate moieties of these enzymes. The phosphomannosyl recognition system is an important mechanism for directing newly synthesized acid hydrolases to lysosomes. The essence of this system is the acquisition of phosphomannosyl residues by acid hydrolases, which leads to binding to specific membrane-bound transport receptors and delivery to lysosomes. However, there is considerable evidence suggesting alternate pathways for targeting acid hydrolases, including alpha-L-fucosidase. This proposal will test the hypothesis that the delivery of newly made alpha-L- fucosidase to lysosomes in lymphoid cells is mediated by phosphomannosyl dependent and independent pathways. The broad objectives are to determine molecular requirements for routing of alpha-L-fucosidase in lymphoid cells and to determine the molecular basis of the defects in the trafficking of alpha-L-fucosidase in lymphoid cell lines derived from fucosidosis and I- cell disease patients. Specific objectives are to research 1) the phosphorylation of alpha-L-fucosidase, 2) the type and content of phosphomannosyl receptors, 3) the subcellular localization of alpha-L- fucosidase, and 4) the precise nature of mutations causing fucosidosis and abnormal secretion of alpha-L-fucosidase. These investigations should provide 1) information concerning requirements for trafficking of alpha-L- fucosidosis and I-cell disease, 3) information about alternative mechanisms for trafficking of acid hydrolases. These experiments could provide a rational basis for treatment of disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK032161-05
Application #
3230615
Study Section
Medical Biochemistry Study Section (MEDB)
Project Start
1985-04-01
Project End
1994-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
5
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Roswell Park Cancer Institute Corp
Department
Type
DUNS #
City
Buffalo
State
NY
Country
United States
Zip Code
14263
Yang, M; DiCioccio, R A (1994) A Gln-281 to Arg substitution in alpha-L-fucosidase is responsible for a common polymorphism detected by isoelectric focusing. Hum Genet 93:597-9
Seo, H C; Yang, M; Tonlorenzi, R et al. (1994) A missense mutation (S63L) in alpha-L-fucosidase is responsible for fucosidosis in an Italian patient. Hum Mol Genet 3:2065-6
Yang, M; Allen, H; DiCioccio, R A (1993) Pedigree analysis of alpha-L-fucosidase gene mutations in a fucosidosis family. Biochim Biophys Acta 1182:245-9
DiCioccio, R A; Miller, A L (1993) Phosphorylation and subcellular location of alpha-L-fucosidase in lymphoid cells from patients with I-cell disease and pseudo-Hurler polydystrophy. Glycobiology 3:489-95
Yang, M; Allen, H; DiCioccio, R A (1992) A mutation generating a stop codon in the alpha-L-fucosidase gene of a fucosidosis patient. Biochem Biophys Res Commun 189:1063-8
Sharma, A; DiCioccio, R A; Allen, H J (1992) Identification and synthesis of a novel 15 kDa beta-galactoside-binding lectin in human leukocytes. Glycobiology 2:285-92
Ahmed, H; Sharma, A; DiCioccio, R A et al. (1992) Lymphoblastoid cell adhesion mediated by a dimeric and polymeric endogenous beta-galactoside-binding lectin (galaptin). J Mol Recognit 5:1-8
Dicioccio, R A; Miller, A L (1992) Binding receptors for alpha-L-fucosidase in human B-lymphoid cell lines. Glycoconj J 9:56-62
DiCioccio, R A; Gordon, B A (1991) Defective expression of alpha-L-fucosidase by lymphoid cells of a fucosidosis patient. Clin Biochem 24:265-70
Allen, H J; Gottstine, S; Sharma, A et al. (1991) Synthesis, isolation, and characterization of endogenous beta-galactoside-binding lectins in human leukocytes. Biochemistry 30:8904-10

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