The role of calmodulin in regulating calcium-permeable PKD2L1 channel activity
10.4196/kjpp.2019.23.3.219
- Author:
Eunice Yon June PARK
1
;
Julia Young BAIK
;
Misun KWAK
;
Insuk SO
Author Information
1. Department of Physiology, Seoul National University College of Medicine, Seoul 03080, Korea. insuk@snu.ac.kr
- Publication Type:Original Article
- Keywords:
Calcium;
Calmodulin;
Ion channel;
Polycystic kidney;
Transient receptor potential channels
- MeSH:
Binding Sites;
Calcium;
Calcium Signaling;
Calmodulin;
Ion Channels;
Ions;
Patch-Clamp Techniques;
Polycystic Kidney Diseases;
Transient Receptor Potential Channels
- From:The Korean Journal of Physiology and Pharmacology
2019;23(3):219-227
- CountryRepublic of Korea
- Language:English
-
Abstract:
Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when coexpressed with CaM and CaMΔN. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ΔEF-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.
