Cytosolic domain regulates the calcium sensitivity and surface expression of BEST1 channels in the HEK293 cells
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BEST family is a class of Ca2+-activated Cl- channels evolutionarywell conserved from bacteria to human. The human BESTparalogs (BEST1-BEST4) share significant amino acid sequencehomology in the N-terminal region, which forms the transmembranehelicases and contains the direct calcium-bindingsite, Ca2+-clasp. But the cytosolic C-terminal region is lessconserved in the paralogs. Interestingly, this domain-specificsequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in theBEST channels is still poorly understood. Thus, we aimed tounderstand the functional role of the C-terminal region in thehuman and mouse BEST1 channels by using electrophysiologicalrecordings. We found that the calcium-dependent activationof BEST1 channels can be modulated by the C-terminalregion. The C-terminal deletion hBEST1 reduced the Ca2+-dependent current activation and the hBEST1-mBEST1 chimerashowed a significantly reduced calcium sensitivity to hBEST1in the HEK293 cells. And the C-terminal domain could regulatecellular expression and plasma membrane targeting ofBEST1 channels. Our results can provide a basis for understandingthe C-terminal roles in the structure-function of BESTfamily proteins.
Abstract
BEST family is a class of Ca2+-activated Cl- channels evolutionarywell conserved from bacteria to human. The human BESTparalogs (BEST1-BEST4) share significant amino acid sequencehomology in the N-terminal region, which forms the transmembranehelicases and contains the direct calcium-bindingsite, Ca2+-clasp. But the cytosolic C-terminal region is lessconserved in the paralogs. Interestingly, this domain-specificsequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in theBEST channels is still poorly understood. Thus, we aimed tounderstand the functional role of the C-terminal region in thehuman and mouse BEST1 channels by using electrophysiologicalrecordings. We found that the calcium-dependent activationof BEST1 channels can be modulated by the C-terminalregion. The C-terminal deletion hBEST1 reduced the Ca2+-dependent current activation and the hBEST1-mBEST1 chimerashowed a significantly reduced calcium sensitivity to hBEST1in the HEK293 cells. And the C-terminal domain could regulatecellular expression and plasma membrane targeting ofBEST1 channels. Our results can provide a basis for understandingthe C-terminal roles in the structure-function of BESTfamily proteins.
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