Calbindin independence of calcium transport in developing teeth contradicts the calcium ferry dogma.
Cytosolic calcium-binding proteins termed calbindins are widely regarded as a key component of the machinery used to transport calcium safely across cells. Acting as mobile buffers, calbindins are thought to ferry calcium in bulk and simultaneously protect against its potentially cytotoxic effects. Here, we contradict this dogma by showing that teeth and bones were produced normally in null mutant mice lacking calbindin(28kDa). Structural analysis of dental enamel, the development of which depends critically on active calcium transport, showed that mineralization was unaffected in calbindin(28kDa)-null mutants. An unchanged rate of calcium transport was verified by measurements of (45)Ca incorporation into developing teeth in vivo. In enamel-forming cells, the absence of calbindin(28kDa) was not compensated by other cytosolic calcium-binding proteins as detectable by (45)Ca overlay, two-dimensional gel, and equilibrium binding analyses. Despite a 33% decrease in cytosolic buffer capacity, cytotoxicity was not evident in either the null mutant enamel or its formative cells. This is the first definitive evidence that calbindins are not required for active calcium transport, either as ferries or as facilitative buffers. Moreover, in challenging the broader notion of a cytosolic route for calcium, the findings support an alternative paradigm involving passage via calcium-tolerant organelles.
Turnbull
Chris I
CI
Departments of Biochemistry and Physiology, University of Otago, PO Box 56, Dunedin 9001, New Zealand.
Looi
Ken
K
Mangum
Jonathan E
JE
Meyer
Michael
M
Sayer
Rod J
RJ
Hubbard
Michael J
MJ
D000818:Animals; D064026:Calbindins; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D010006:Osteoblasts / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D064030:S100 Calcium Binding Protein G / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D013997:Time Factors; D014070:Tooth / Q000196:embryology*
D064026:Calbindins; D064030:S100 Calcium Binding Protein G; D002118:Calcium