Binding of a dihydropyridine felodipine-analogue to calmodulin and related calcium-binding proteins.
A dihydropyridine-affinity column was prepared by coupling a physiologically active and vasoselective amino-derivative of felodipine to divinylsulfone-activated Trisacryl GF2000. Calmodulin (CaM) as well as the homologous calcium-binding proteins skeletal and cardiac Troponin C (sTnC and cTnC) and S100b bound to this resin in a calcium-dependent manner. In contrast, other homologous proteins such as parvalbumin and the intestinal calcium-binding protein did not bind. Competition studies showed that CaM had a higher affinity for the felodipine-column than sTnC or cTnC. Through studies with a series of proteolytic fragments of CaM and sTnC, it was found that the felodipine binding site is located in the amino-terminal domain of the protein. These results illustrate the utility of affinity-chromatography for the study of dihydropyridine-binding proteins.
Boström
S L
SL
Department of Cardiovascular Pharmacology, Hassle Research Laboratories, Mölndal, Sweden.
Westerlund
C
C
Rochester
S
S
Vogel
H J
HJ
D001667:Binding, Competitive; D002121:Calcium Channel Blockers / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002846:Chromatography, Affinity; D015736:Felodipine; D009568:Nitrendipine / Q000031:analogs & derivatives* / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D013329:Structure-Activity Relationship; D014336:Troponin / Q000378:metabolism; D019209:Troponin C
D002121:Calcium Channel Blockers; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D010446:Peptide Fragments; D014336:Troponin; D019209:Troponin C; D009568:Nitrendipine; D015736:Felodipine