The proposed studies are designed to achieve a systematic and comprehensive analysis of the multigene family of salivary kallikreins in the rat and human. This analysis will focus upon gene expression and the regulation and distribution of salivary kallikreins at the cellular and subcellular levels. Tissue kallikreins are closely related serine proteases which specifically cleave peptide precursors to release biologically active peptides. Although kallikreins are found in many diverse tissues, their regulation and expression in salivary glands are important because of their potential role in processing a wide variety of substrates which may be of major importance in the maintenance of oral health. Recently developed tools have allowed us to sequence the complete rat kallikrein gene, the tonin gene, and to partially sequence several additional kallikrein-related genes. These sequences have provided valuable oligonucleotide probes to study the regulation of this family of enzymes. Further definition of the role of tissue kallikrein-related enzymes in salivary glands will be approached by: 1) complete mapping and sequencing of the cDNA for rat and human salivary kallikrein and other kallikrein gene family members, which will enable specific probe design for identifying and quantifying salivary kallikrein gene transcripts; 2) analyzing salivary kallikrein expression in vitro by deletion mapping, gel retardation assays and foot-printing techniques; 3) purifying and characterizing human and rat kallikreins to gain insight into their enzymatic properties and substrate specificities and to generate polyclonal and monoclonal antibodies for their individual identification and quantification; 4) elucidating the hormonal regulation of rat salivary kallikrein gene expression by specific RIA, Northern blotting, in situ hybridization, and immunocytochemical methods; 5) evaluating the subcellular compartmentalization of salivary kallikreins to gain insight into the sorting and secretory mechanisms of individual kallikrein gene family members.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE009731-04
Application #
2130720
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1991-01-01
Project End
1995-12-31
Budget Start
1994-01-01
Budget End
1994-12-31
Support Year
4
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Medical University of South Carolina
Department
Biochemistry
Type
Schools of Medicine
DUNS #
183710748
City
Charleston
State
SC
Country
United States
Zip Code
29425
Richards, G P; Zintz, C; Chao, J et al. (1996) Purification and characterization of salivary kallikrein from an insectivore (Scalopus aquaticus): substrate specificities, immunoreactivity, and kinetic analyses. Arch Biochem Biophys 329:104-12
Yu, J X; Chao, L; Ward, D C et al. (1996) Structure and chromosomal localization of the human prostasin (PRSS8) gene. Genomics 32:334-40
Chai, K X; Ni, A; Wang, D et al. (1996) Genomic DNA sequence, expression, and chromosomal localization of the human B1 bradykinin receptor gene BDKRB1. Genomics 31:51-7
Richards, G P; Chao, J; Chao, L (1996) Tissue kallikreins in evolutionarily diverse vertebrates. Immunopharmacology 32:94-5
Chao, J; Schmaier, A; Chen, L M et al. (1996) Kallistatin, a novel human tissue kallikrein inhibitor: levels in body fluids, blood cells, and tissues in health and disease. J Lab Clin Med 127:612-20
Garrett, J R; Chao, J; Proctor, G B et al. (1995) Influences of secretory activities in rat submandibular glands on tissue kallikrein circulating in the blood. Exp Physiol 80:429-40
Lin, K F; Chao, J; Chao, L (1995) Human atrial natriuretic peptide gene delivery reduces blood pressure in hypertensive rats. Hypertension 26:847-53
Yu, J X; Chao, L; Chao, J (1995) Molecular cloning, tissue-specific expression, and cellular localization of human prostasin mRNA. J Biol Chem 270:13483-9
Simson, J A; Chao, J (1994) Subcellular distribution of tissue kallikrein and Na,K-ATPase alpha-subunit in rat parotid striated duct cells. Cell Tissue Res 275:407-17
Simson, J A; Wang, J; Chao, J et al. (1994) Histopathology of lymphatic tissues in transgenic mice expressing human tissue kallikrein gene. Lab Invest 71:680-7

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