title: Phosphatidylinositol 3-phosphate indirectly activates KCa3.1 via 14 amino acids in the carboxy terminus of KCa3.1.
abstract: KCa3.1 is an intermediate conductance Ca2+-activated K+ channel that is expressed predominantly in hematopoietic cells, smooth muscle cells, and epithelia where it functions to regulate membrane potential, Ca2+ influx, cell volume, and chloride secretion. We recently found that the KCa3.1 channel also specifically requires phosphatidylinositol-3 phosphate [PI(3)P] for channel activity and is inhibited by myotubularin-related protein 6 (MTMR6), a PI(3)P phosphatase. We now show that PI(3)P indirectly activates KCa3.1. Unlike KCa3.1 channels, the related KCa2.1, KCa2.2, or KCa2.3 channels do not require PI(3)P for activity, suggesting that the KCa3.1 channel has evolved a unique means of regulation that is critical for its biological function. By making chimeric channels between KCa3.1 and KCa2.3, we identified a stretch of 14 amino acids in the carboxy-terminal calmodulin binding domain of KCa3.1 that is sufficient to confer regulation of KCa2.3 by PI(3)P. However, mutation of a single potential phosphorylation site in these 14 amino acids did not affect channel activity. These data together suggest that PI(3)P and these 14 amino acids regulate KCa3.1 channel activity by recruiting an as yet to be defined regulatory subunit that is required for Ca2+ gating of KCa3.1.
authors.affiliation: Department of Pharmacology, The Skirball Institute, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Shekhar
authors.initials: S
authors.lastname: Srivastava
authors.forename: Papiya
authors.initials: P
authors.lastname: Choudhury
authors.forename: Zhai
authors.initials: Z
authors.lastname: Li
authors.forename: Gongxin
authors.initials: G
authors.lastname: Liu
authors.forename: Vivek
authors.initials: V
authors.lastname: Nadkarni
authors.forename: Kyung
authors.initials: K
authors.lastname: Ko
authors.forename: William A
authors.initials: WA
authors.lastname: Coetzee
authors.forename: Edward Y
authors.initials: EY
authors.lastname: Skolnik
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D017124:Conserved Sequence; D006224:Cricetinae; D003600:Cytosol; D004594:Electrophysiology; D004789:Enzyme Activation; D006801:Humans; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D018408:Patch-Clamp Techniques; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases; D010766:Phosphorylation; D054558:Protein Tyrosine Phosphatases, Non-Receptor; D051381:Rats; D016415:Sequence Alignment
chemical_list: D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C496923:KCNN4 protein, human; D010446:Peptide Fragments; D018129:Phosphatidylinositol Phosphates; C055525:phosphatidylinositol 3-phosphate; D010744:Phosphoric Monoester Hydrolases; C513490:MTMR6 protein, human; D054558:Protein Tyrosine Phosphatases, Non-Receptor