FKBP-12 exhibits an inhibitory activity on calcium oxalate crystal growth in vitro.
Urolithiasis and calcium oxalate crystal deposition diseases are still significant medical problems. In the course of nephrocalcin cDNA cloning, we have identified FKBP-12 as an inhibitory molecule of calcium oxalate crystal growth. lambdagt 11 cDNA libraries were constructed from renal carcinoma tissues and screened for nephrocalcin cDNA clones using anti-nephrocalcin antibody as a probe. Clones expressing recombinant proteins, which appeared to be antigenically cross-reactive to nephrocalcin, were isolated and their DNA sequences and inhibitory activities on the calcium oxalate crystal growth were determined. One of the clone lambda gt 11 #31-1 had a partial fragment (80 bp) of FKBP-12 cDNA as an insert. Therefore, a full-length FKBP-12 cDNA was PCR-cloned from the lambda gt 11 renal carcinoma cDNA library and was subcloned into an expression vector. The resultant recombinant FKBP-12 exhibited an inhibitory activity on the calcium oxalate crystal growth (Kd=10(-7) M). Physiological effect of the extracellular FKBP-12 was investigated in terms of macrophage activation and proinflammatory cytokine gene induction. Extracellular FKBP-12 failed to activate macrophages even at high concentrations. FKBP-12 seems an anti-stone molecule for the oxalate crystal deposition disease and recurrent stone diseases.
Han
In Sook
IS
Department of Immunology, Institute for Medical Science, Keimyung University, Taegu, Korea.
Nakagawa
Yasushi
Y
Park
Jong Wook
JW
Suh
Min Ho
MH
Suh
Sung Il
SI
Shin
Song Woo
SW
Ahn
Su Yul
SY
Choe
Byung Kil
BK
D000818:Animals; D001483:Base Sequence; D002129:Calcium Oxalate / Q000037:antagonists & inhibitors*; D002292:Carcinoma, Renal Cell; D003460:Crystallization; D018076:DNA, Complementary; D005110:Extracellular Space; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D007669:Kidney Calculi / Q000517:prevention & control*; D007680:Kidney Neoplasms; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D022061:Tacrolimus Binding Protein 1A / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins; C029490:nephrocalcin; D002129:Calcium Oxalate; D022061:Tacrolimus Binding Protein 1A