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J Cell Sci


Title:Redox regulation of the yeast voltage-gated Ca(2+) channel homolog Cch1p by glutathionylation of specific cysteine residues
Author(s):Chandel A; Bachhawat AK;
Address:"Department of Biological Sciences, Indian Institute of Science Education & Research (IISER), Sector 81, Mohali, Punjab 140306, India. Department of Biological Sciences, Indian Institute of Science Education & Research (IISER), Sector 81, Mohali, Punjab 140306, India anand@iisermohali.ac.in"
Journal Title:J Cell Sci
Year:2017
Volume:20170602
Issue:14
Page Number:2317 - 2328
DOI: 10.1242/jcs.202853
ISSN/ISBN:1477-9137 (Electronic) 0021-9533 (Linking)
Abstract:"Cch1p, the yeast homolog of the pore-forming subunit alpha(1) of the mammalian voltage-gated Ca(2+) channel (VGCC), is located on the plasma membrane and mediates the redox-dependent influx of Ca(2+) Cch1p is known to undergo both rapid activation (after oxidative stress and or a change to high pH) and slow activation (after ER stress and mating pheromone activation), but the mechanism of activation is not known. We demonstrate here that both the fast activation (exposure to pH 8-8.5 or treatment with H(2)O(2)) and the slow activation (treatment with tunicamycin or alpha-factor) are mediated through a common redox-dependent mechanism. Furthermore, through mutational analysis of all 18 exposed cysteine residues in the Cch1p protein, we show that the four mutants C587A, C606A, C636A and C642A, which are clustered together in a common cytoplasmic loop region, were functionally defective for both fast and slow activations, and also showed reduced glutathionylation. These four cysteine residues are also conserved across phyla, suggesting a conserved mechanism of activation. Investigations into the enzymes involved in the activation reveal that the yeast glutathione S-transferase Gtt1p is involved in the glutathionylation of Cch1p, while the thioredoxin Trx2p plays a role in the Cch1p deglutathionylation"
Keywords:Alanine/genetics Amino Acid Sequence Calcium Channels/genetics/*metabolism Conserved Sequence Cysteine/genetics/*metabolism Cytoplasm/metabolism Glutathione/*metabolism Hydrogen-Ion Concentration Mutation Oxidation-Reduction Oxidative Stress/physiology Sa;
Notes:"MedlineChandel, Avinash Bachhawat, Anand K eng England 2017/06/04 J Cell Sci. 2017 Jul 15; 130(14):2317-2328. doi: 10.1242/jcs.202853. Epub 2017 Jun 2"

 
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