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CRAC channel is inhibited by neomycin in a Ptdlns(4,5)P2‐independent manner
Author(s) -
Huang Kun,
Wang Xuemei,
Liu Yanjun,
Zhao Yi
Publication year - 2015
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.3088
Subject(s) - neomycin , orai1 , phosphatidylinositol , chemistry , microbiology and biotechnology , patch clamp , biophysics , intracellular , phosphatase , stim1 , biochemistry , membrane , biology , kinase , phosphorylation , receptor , antibiotics
Depletion of intracellular Ca 2+ stores evokes store‐operated Ca 2+ entry through the Ca 2+ release‐activated Ca 2+ (CRAC) channels. In this study, we found that the store‐operated Ca 2+ entry was inhibited by neomycin, an aminoglycoside that strongly binds phosphatidylinositol 4,5‐bisphosphate (PtdIns(4,5)P2). Patch clamp recordings revealed that neomycin blocked the CRAC currents reconstituted by co‐expression of Orai1 and Stim1 in HEK293 cells. Using a rapamycin‐inducible PtdIns(4,5)P2‐specific phosphatase (Inp54p) system to manipulate the PtdIns(4,5)P2 in the plasma membrane, we found that the CRAC current was not altered by PtdIns(4,5)P2 depletion. This result suggests that PtdIns(4,5)P2 is not required for CRAC channel activity, and thereby, neomycin inhibits CRAC channels in a manner that is independent of neomycin–PtdIns(4,5)P2 binding. Copyright © 2015 John Wiley & Sons, Ltd.