Effects of temperature on human L-type cardiac Ca2+ channels expressed in Xenopus oocytes.
Temperature normally affects peak L-type Ca2+ channel (CaCh) current with a temperature coefficient (Q10) of between 1.8 and 3.5; in cardiomyocytes attenuating protein kinase A activity increases Q10 whilst activating it lowers Q10. We examine temperature effects using cloned human cardiac CaChs expressed in Xenopus oocytes. Peak inward currents (IBa) through expressed CaChs (i.e. alpha1C alpha2/deltaa beta1b) exhibited a Q10 of 5.8+/-0.4 when examined between 15 and 25 degreesC. The nifedipine-sensitive IBa exhibited a higher Q10 of 8.7+/-0.5, whilst the nifedipine-insensitive IBa exhibited Q10 of 3.7+/-0.3. Current/voltage (I/V) relationships shifted to negative potentials on warming. Using instead a different CaCh beta subunit isoform, beta2c, gave rise to an IBa similar to those expressed using beta1b. We utilized a carboxyl deletion mutant, alpha1C-Delta1633, to determine the temperature sensitivity of the pore moiety in the absence of auxiliary subunits; IBa through this channel exhibited a Q10 of 9.3+/-0.3. However, the Q10 for macroscopic conductance was reduced compared to that of heteromeric channels; decreasing from 5.0 (i.e. alpha1C alpha2/deltaa beta1b) and 3.9 (i.e. alpha1C alpha2/deltaa beta2c) to 2.4 (alpha1C-Delta1633). These observations differ markedly from those made in studies of cardiomyocytes, and suggest that enhanced sensitivity may depend on the membrane environment, channel assembly or other regulatory factors.
Allen
T J
TJ
Department of Pharmacology, Royal Free Hospital School of Medicine, London, NW3 2PF, UK.
Mikala
G
G
D000595:Amino Acid Sequence; D000818:Animals; D015220:Calcium Channels / Q000235:genetics* / Q000502:physiology*; D020746:Calcium Channels, L-Type; D004553:Electric Conductivity; D005260:Female; D017353:Gene Deletion; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009206:Myocardium / Q000737:chemistry; D009865:Oocytes / Q000378:metabolism*; D011994:Recombinant Proteins; D016415:Sequence Alignment; D013696:Temperature; D014982:Xenopus laevis
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D011994:Recombinant Proteins