Cyclosporine A decreases the protein level of the calcium-binding protein calbindin-D 28kDa in rat kidney.
Despite the widespread use of cyclosporine A (CsA), its mechanism of action and side effects are not yet completely understood. There exists a large body of evidence suggesting that disturbance of calcium homeostasis is a critical step in the cascade of cellular and molecular events induced by the drug. As recently shown in our laboratory by two-dimensional protein gel electrophoresis (2-DE) analysis of kidney homogenates, CsA induced numerous changes in several kidney proteins. One kidney protein in particular was shown to be strongly down-regulated by the drug. In this work we report the identification of the strongly decreased kidney protein as calbindin-D 28kDa, a vitamin D-dependent calcium-binding protein associated with calcium handling by cells. The assignment of the down-regulated protein spot is based on its internal amino acid sequence analysis and its specific reaction with a monoclonal antibody raised against calbindin-D 28kDa. In kidney homogenates of male Wistar rats treated with 50 mg/kg/d CsA for up to 28 days, calbindin levels were measured by ELISA and were shown to be continuously decreased with prolonged CsA treatment. To our knowledge, this is the first report describing the effect of CsA on kidney calbindin-D 28kDa protein levels. Further studies are needed to elucidate whether the CsA-mediated down-regulation of the calcium-binding protein calbindin-D 28kDa may be a critical factor for the renal adverse effects induced by this drug.
Steiner
S
S
Drug Safety Assessment, Toxicology, Sandoz Pharma Ltd., Basel, Switzerland.
Aicher
L
L
Raymackers
J
J
Meheus
L
L
Esquer-Blasco
R
R
Anderson
N L
NL
Cordier
A
A
D000595:Amino Acid Sequence; D000818:Animals; D064026:Calbindins; D016572:Cyclosporine / Q000494:pharmacology*; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D051381:Rats; D017208:Rats, Wistar; D064030:S100 Calcium Binding Protein G / Q000378:metabolism*
D064026:Calbindins; D007166:Immunosuppressive Agents; D064030:S100 Calcium Binding Protein G; D016572:Cyclosporine