The overall objective of this research program is to elucidate the mechanisms which regulate the expression of proteoglycans by focussing on the structure and synthesis of the prototype proteoglycan of cartilage. These extracellular macromolecules, along with hyaluronic acid and collagen, exhibit changing patterns in expression during development of cartilage tissue. As well, alterations n these components may underlie certain disorders of cartilage resulting in abnormal growth, development or function. Thus understanding the mechanisms which control their expression is a fundamental problem in connective tissue biochemistry. Specific questions to be addressed and hypothesis to be tested include: 1) elucidating the role of specific amino acid sequences (recognition or consensus) in marking sites of carbohydrate substitutions in core proteins, 2) exploring the structural organization of core protein domains and the relatedness of core protein families expressed by different cells and tissues, 3) defining the mechanisms by which the intracellular processing machinery is organized and controlled 4) establishing the significance of gene structure to the diversity and ontogeny of the characteristic domain organization of proteoglycans, 5) defining the mechanism of action of specific enzymes responsible for initiating and processing proteoglycans. These will be accomplished by a combinational approach using direct amino acid sequencing of carbohydrate-substituted peptides, nucleotide sequencing of cDNA clones coding for these regions of core proteins and synthesizing artificial peptides to be tested as model acceptors. Biosynthetic studies will take advantage of a newly developed method for preparing permeabilized cells that can be labeled directly with nucleotide sugar precursors. The mechanism of action of relevant enzymes will be examined using appropriate peptide fragments and newly synthesized substrate analogs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AR019622-23
Application #
6043190
Study Section
Special Emphasis Panel (NSS)
Program Officer
Tyree, Bernadette
Project Start
1977-08-15
Project End
2001-07-31
Budget Start
1999-08-01
Budget End
2001-07-31
Support Year
23
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Chicago
Department
Pediatrics
Type
Schools of Medicine
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
Domowicz, Miriam S; Mueller, Melissa M; Novak, Todd E et al. (2003) Developmental expression of the HNK-1 carbohydrate epitope on aggrecan during chondrogenesis. Dev Dyn 226:42-50
Schwartz, Nancy B; Domowicz, Miriam (2002) Chondrodysplasias due to proteoglycan defects. Glycobiology 12:57R-68R
Pirok 3rd, E W; Henry, J; Schwartz, N B (2001) cis elements that control the expression of chick aggrecan. J Biol Chem 276:16894-903
Domowicz, M S; Pirok 3rd, E W; Novak, T E et al. (2000) Role of the C-terminal G3 domain in sorting and secretion of aggrecan core protein and ubiquitin-mediated degradation of accumulated mutant precursors. J Biol Chem 275:35098-105
Schwartz, N (2000) Biosynthesis and regulation of expression of proteoglycans. Front Biosci 5:D649-55
Krueger Jr, R C; Kurima, K; Schwartz, N B (1999) Completion of the mouse aggrecan gene structure and identification of the defect in the cmd-Bc mouse as a near complete deletion of the murine aggrecan gene. Mamm Genome 10:1119-25
Kurima, K; Singh, B; Schwartz, N B (1999) Genomic organization of the mouse and human genes encoding the ATP sulfurylase/adenosine 5'-phosphosulfate kinase isoform SK2. J Biol Chem 274:33306-12
Immergluck, L C; Domowicz, M S; Schwartz, N B et al. (1998) Viral and cellular requirements for entry of herpes simplex virus type 1 into primary neuronal cells. J Gen Virol 79 ( Pt 3):549-59
Schwartz, N B; Lyle, S; Ozeran, J D et al. (1998) Sulfate activation and transport in mammals: system components and mechanisms. Chem Biol Interact 109:143-51
Pirok 3rd, E W; Li, H; Mensch Jr, J R et al. (1997) Structural and functional analysis of the chick chondroitin sulfate proteoglycan (aggrecan) promoter and enhancer region. J Biol Chem 272:11566-74

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