Heparin binds with high affinity to voltage-dependent L-type Ca2+ channels. Evidence for an agonistic action.
Heparin and related polyanions are a new class of compounds interacting with 1,4-dihydropyridine-sensitive L-type Ca2+ channels in a tissue-specific manner. Labeling of membrane-bound Ca2+ channels in rabbit skeletal muscle transverse tubules at the phenylalkylamine, benzothiazepine, and 1,4-dihydropyridine-selective domains was inhibited reversibly by a noncompetitive mechanism as shown by equilibrium saturation analysis and kinetic studies. (+)-cis-diltiazem but not (-)-cis-diltiazem reduced the inhibitory potency of heparin for 1,4-dihydropyridines. Antagonistic but not agonistic 1,4-dihydropyridines reversed heparin inhibition at the benzothiazepine site. Heparin forms a tight complex with the purified Ca2+ channel which is highly sensitive with respect to heparin inhibition (IC50 value: 0.05 microgram/ml) of 1,4-dihydropyridine binding. Reconstituted channel complexes have completely lost 1,4-dihydropyridine binding-inhibition by heparin and are not retained by lectin or heparin affinity columns. In whole cell patch clamp experiments with guinea-pig cardiac myocytes heparin increased the current through L-type Ca2+ channels when applied extracellulary. Synthetic peptides (representing putative heparin binding domains) which were derived from the rabbit skeletal muscle alpha 1-subunit reversed the inhibitory effects of heparin on 1,4-dihydropyridine receptors. Reversal for a peptide representing an extracellular domain occurred by an apparently competitive mechanism. It is suggested that heparin and related polyanions may interact with an evolutionary conserved cluster of basic amino acids in the large putative extracellular domain connecting the fifth and sixth putative transmembrane segment in the first motif of the ionic pore-forming alpha 1-subunit from skeletal muscle.
Knaus
H G
HG
Institute for Biochemical Pharmacology, Innsbruck, Austria.
Scheffauer
F
F
Romanin
C
C
Schindler
H G
HG
Glossmann
H
H
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D015220:Calcium Channels / Q000378:metabolism*; D002347:Carps; D002462:Cell Membrane / Q000378:metabolism; D002540:Cerebral Cortex / Q000378:metabolism; D004095:Dihydropyridines / Q000037:antagonists & inhibitors / Q000378:metabolism; D004110:Diltiazem / Q000378:metabolism / Q000494:pharmacology; D004553:Electric Conductivity; D006168:Guinea Pigs; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D017275:Isradipine; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D018733:Nicotinic Antagonists; D010069:Oxadiazoles / Q000378:metabolism; D000071228:Polyelectrolytes; D011108:Polymers / Q000378:metabolism; D011817:Rabbits; D011978:Receptors, Nicotinic / Q000378:metabolism; D013237:Stereoisomerism; D014700:Verapamil / Q000031:analogs & derivatives / Q000378:metabolism
D015220:Calcium Channels; D004095:Dihydropyridines; D004110:Diltiazem; D006493:Heparin; D017275:Isradipine; D018733:Nicotinic Antagonists; D010069:Oxadiazoles; D000071228:Polyelectrolytes; D011108:Polymers; D011978:Receptors, Nicotinic; D014700:Verapamil; C038806:1,4-dihydropyridine; C043184:4-desmethoxyverapamil