The presence and regulation of Ca-binding proteins (CaBPs) were investigated in newly identified 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] target tissues. 45Ca(2+)-blot analysis of proteins in normal rats yielded a 45Ca2+ band comigrating with authentic calmodulin. Additionally, a parvalbumin-like band (mol mass = 15.4 +/- 0.3 kDa) was prominent in prostate, and a strong unidentified 45Ca2+ band was always evident in the testis (mol mass = 23.5 +/- 0.7 kDa). Lung, bladder, and especially prostate demonstrated 45Ca2+ bands comigrating with the intestinal vitamin D-related CaBP (CaBP-D9K; mol mass = 10.9 +/- 0.5 kDa). Most tissues (including testis, heart, and lung) exhibited low levels of a 45Ca2+ band comigrating with the renal CaBP-D28K (mol mass = 28.3 +/- 0.4 kDa). Importantly, 45Ca2+ binding to all detectable CaBPs was unchanged in these four tissues in vitamin D-deficient rats, despite substantial downregulation of the intestinal CaBP-D9K and renal CaBP-D28K. Neither immunoblot analysis (rabbit anti-rat renal CaBP-D28K) nor Northern analysis (rat brain CaBP-D28K cDNA) provided evidence for coidentity of the 28-kDa 45Ca2+ band with the CaBP-D28K. Conversely, immunoblot analysis of lung, but not prostate, cytosol provided evidence for specific immunocross-reactivity to rabbit anti-rat intestinal CaBP-D9K. Immunoblot analysis of the 9-kDa CaBP in lung further confirmed its vitamin D independence. In conclusion, the vitamin D independence of the CaBPs in these putative new 1,25(OH)2D3 targets suggests the absence of an obligatory relationship between 1,25(OH)2D3 effects and CaBP induction therein.(ABSTRACT TRUNCATED AT 250 WORDS)
The Molecular Cloning of the Complementary Deoxyribonucleic Acid for Bovine Vitamin D-Dependent Calcium-Binding Protein: Structure of the Full-Length Protein and Evidence for Homologies with Other Calcium-Binding Proteins of the Troponin-C Superfamily of Proteins
