A fluoride-sensitive acid phosphatase activity is clearly present in osteoblast-line cells, however, its function is unknown. Based on the preliminary findings regarding this enzyme which are described in this application, the following hypotheses have been developed: (1) F-sensitive acid phosphatase is unique to osteoblast-line cells, (2) this enzyme acts as a neutral pH phosphotyrosyl phosphatase in proliferating bone cells, (3) this enzyme is, at least in part, a significant determinant of phosphotyrosyl protein levels in these cells, and (4) inhibition of this enzyme by specific inhibitors will serve to increase tyrosyl phosphorylation and stimulate bone cell proliferation. In this application, the first two hypotheses will be tested by purifying and characterizing this enzyme (and its neutral pH phosphotyrosyl phosphatase activity) from chick osteoblast-line cells, and determining its primary sequence. Specific antibodies against the purified enzyme will be developed and characterized, and they will be used in studies to test hypotheses 3 and 4. Because it is essential to identify physiologic substrates for an enzyme before its function can be tested, cellular substrates of this F-sensitive acid phosphatase will be identified by determining those cellular proteins whose tyrosyl phosphorylation are stimulated by (1) inhibitors of the enzyme (ie., F, vanadate, and molybdate) and (2) by specific inhibitory antibodies. [The P-tyr proteins will be isolated with anti-Tyr(P) antibodies.] The long term goal of these studies is to test the model that inhibition of phosphotyrosyl phosphatase can lead to cell proliferation in general (i.e., hypothesis 4). If my hypotheses are correct, the significance of these proposed studies will be two-fold: (1) They would provide important information on a key regulatory process (i.e., cell proliferation) in bone. More importantly, this information I derive about this system of P-tyr phosphorylation may be applicable to all renewing cell populations, not merely bone cells. (2) They will advance our understanding of the molecular mechanism of action of F, which is an effective therapeutic agent in the treatment of spinal osteoporosis. Knowledge of the mechanism of action of F could provide background information for development of new therapeutic treatments, similar in mechanism to F but more effective and less toxic.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE008681-03
Application #
2130145
Study Section
Orthopedics and Musculoskeletal Study Section (ORTH)
Project Start
1990-03-01
Project End
1993-02-28
Budget Start
1992-03-01
Budget End
1993-02-28
Support Year
3
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Loma Linda University
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
City
Loma Linda
State
CA
Country
United States
Zip Code
92350
Lau, K-H William; Baylink, David J (2003) Osteoblastic tartrate-resistant acid phosphatase: its potential role in the molecular mechanism of osteogenic action of fluoride. J Bone Miner Res 18:1897-900
Lau, K-H William; Goodwin, C; Arias, M et al. (2002) Bone cell mitogenic action of fluoroaluminate and aluminum fluoride but not that of sodium fluoride involves upregulation of the insulin-like growth factor system. Bone 30:705-11
Hashimoto, H; Lau, K H (2001) Differential effects of bacterial toxins on mitogenic actions of sodium fluoride and those of aluminum fluoride in human TE85 osteosarcoma cells. Mol Cell Biochem 228:91-8
Yoon, H K; Baylink, D J; Lau, K H (2000) Protein tyrosine kinase inhibitors block the stimulatory actions of phosphotyrosine phosphatase inhibitors to increase cell proliferation, alkaline phosphatase activity, and collagen synthesis in normal human bone cells. Am J Nephrol 20:153-62
Lau, K H; Baylink, D J (1999) Vitamin D therapy of osteoporosis: plain vitamin D therapy versus active vitamin D analog (D-hormone) therapy. Calcif Tissue Int 65:295-306
Lau, K H; Baylink, D J (1998) Molecular mechanism of action of fluoride on bone cells. J Bone Miner Res 13:1660-7
Yoon, H K; Chen, K; Baylink, D J et al. (1998) Differential effects of two protein tyrosine kinase inhibitors, tyrphostin and genistein, on human bone cell proliferation as compared with differentiation. Calcif Tissue Int 63:243-9
Yoo, A; Tanimoto, H; Akesson, K et al. (1998) Effects of calcium depletion and repletion on serum insulin-like growth factor I and binding protein levels in weanling rats. Bone 22:225-32
Dimai, H P; Porta, S; Wirnsberger, G et al. (1998) Daily oral magnesium supplementation suppresses bone turnover in young adult males. J Clin Endocrinol Metab 83:2742-8
Akesson, K; Lau, K H; Johnston, P et al. (1998) Effects of short-term calcium depletion and repletion on biochemical markers of bone turnover in young adult women. J Clin Endocrinol Metab 83:1921-7

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