title: Lipids modulate the increase of BK channel calcium sensitivity by the β1 subunit.
abstract: Co-expression of the auxiliary β1 subunit with the pore forming α subunit of BK dramatically alters apparent calcium sensitivity. Investigation of the mechanism underlying the increase in calcium sensitivity of BK in smooth muscle has concentrated on the energetic effect of β1's interaction with α. We take a novel approach, exploring whether β1 modification of calcium sensitivity reflects altered interaction between the channel protein and surrounding lipids. We reconstituted hSlo BK α and BK α+β1 channels into two sets of bilayers. One set contained POPE with POPS, POPG, POPA and POPC, where the length of acyl chains is constant, but surface charge differs. The second set is a series of neutral bilayers formed from DOPE with phosphatidylcholines (PCs) of varying acyl chain lengths: C (14:1), C (18:1), C (22:1) and C (24:1), and with brain sphingomyelin (SPM), in which surface charge is constant, but bilayer thickness varies. The increase in calcium sensitivity caused by the β1 subunit was preserved in negatively charged lipid bilayers but not in neutral bilayers, indicating that modification of apparent Ca(2+) sensitivity by β1 is modulated by membrane lipids, requiring negatively charged lipids in the membrane. Moreover, the presence of β1 reduces BK activity in thin bilayers of PC 14:1 and thick bilayers containing SPM, but has no significant effect on activity of BK in PC 18:1, PC 22:1 and PC 24:1 bilayers. These data suggest that auxiliary β1 subunits fine-tune channel gating not only through direct subunit-subunit interactions but also by modulating lipid-protein interactions.
authors.affiliation: Institute of Neurobiology, University of Puerto Rico, San Juan, Puerto Rico.
authors.forename: Chunbo
authors.initials: C
authors.lastname: Yuan
authors.affiliation: Institute of Neurobiology, University of Puerto Rico, San Juan, Puerto Rico.
authors.forename: Cristina
authors.initials: C
authors.lastname: Velázquez-Marrero
authors.affiliation: Institute of Neurobiology, University of Puerto Rico, San Juan, Puerto Rico.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bernardo
authors.affiliation: Institute of Neurobiology, University of Puerto Rico, San Juan, Puerto Rico.
authors.forename: Steven N
authors.initials: SN
authors.lastname: Treistman
mesh_terms: D002118:Calcium / Q000494:pharmacology*; D055724:Electrophysiological Phenomena; D057809:HEK293 Cells; D006801:Humans; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism
chemical_list: D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D002118:Calcium