Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes
Sodium balance is maintained by the precise regulation of the activity of the epithelial sodium channel (ENaC) in the kidney. We have recently reported an extracellular activation of ENaC-mediated sodium transport (I(Na)) by a GPI-anchored serine protease (mouse channel-activating protein, mCAP1) th...
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description | Sodium balance is maintained by the precise regulation of the activity of the epithelial sodium channel (ENaC) in the kidney. We have recently reported an extracellular activation of ENaC-mediated sodium transport (I(Na)) by a GPI-anchored serine protease (mouse channel-activating protein, mCAP1) that was isolated from a cortical collecting duct cell line derived from mouse kidney. In the present study, we have identified two additional membrane-bound serine proteases (mCAP2 and mCAP3) that are expressed in the same cell line. We show that each of these proteases is able to increase I(Na) 6-10-fold in the Xenopus oocyte expression system. I(Na) and the number (N) of channels expressed at the cell surface (measured by binding of a FLAG monoclonal I(125)-radioiodinated antibody) were measured in the same oocyte. Using this assay, we show that mCAP1 increases I(Na) 10-fold (P < 0.001) but N remained unchanged (P = 0.9), indicating that mCAP1 regulates ENaC activity by increasing its average open probability of the whole cell (wcP(o)). The serum- and glucocorticoid-regulated kinase (Sgk1) involved in the aldosterone-dependent signaling cascade enhances I(Na) by 2.5-fold (P < 0.001) and N by 1.6-fold (P < 0.001), indicating a dual effect on N and wcP(o). Compared with Sgk1 alone, coexpression of Sgk1 with mCAP1 leads to a ninefold increase in I(Na) (P < 0.001) and 1.3-fold in N (P < 0.02). Similar results were observed for mCAP2 and mCAP3. The synergism between CAPs and Sgk1 on I(Na) was always more than additive, indicating a true potentiation. The synergistic effect of the two activation pathways allows a large dynamic range for ENaC-mediated sodium regulation crucial for a tight control of sodium homeostasis. |
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We have recently reported an extracellular activation of ENaC-mediated sodium transport (I(Na)) by a GPI-anchored serine protease (mouse channel-activating protein, mCAP1) that was isolated from a cortical collecting duct cell line derived from mouse kidney. In the present study, we have identified two additional membrane-bound serine proteases (mCAP2 and mCAP3) that are expressed in the same cell line. We show that each of these proteases is able to increase I(Na) 6-10-fold in the Xenopus oocyte expression system. I(Na) and the number (N) of channels expressed at the cell surface (measured by binding of a FLAG monoclonal I(125)-radioiodinated antibody) were measured in the same oocyte. Using this assay, we show that mCAP1 increases I(Na) 10-fold (P < 0.001) but N remained unchanged (P = 0.9), indicating that mCAP1 regulates ENaC activity by increasing its average open probability of the whole cell (wcP(o)). The serum- and glucocorticoid-regulated kinase (Sgk1) involved in the aldosterone-dependent signaling cascade enhances I(Na) by 2.5-fold (P < 0.001) and N by 1.6-fold (P < 0.001), indicating a dual effect on N and wcP(o). Compared with Sgk1 alone, coexpression of Sgk1 with mCAP1 leads to a ninefold increase in I(Na) (P < 0.001) and 1.3-fold in N (P < 0.02). Similar results were observed for mCAP2 and mCAP3. The synergism between CAPs and Sgk1 on I(Na) was always more than additive, indicating a true potentiation. The synergistic effect of the two activation pathways allows a large dynamic range for ENaC-mediated sodium regulation crucial for a tight control of sodium homeostasis.</description><identifier>ISSN: 0022-1295</identifier><identifier>EISSN: 1540-7748</identifier><identifier>DOI: 10.1085/jgp.20028598</identifier><identifier>PMID: 12149280</identifier><language>eng</language><publisher>United States: The Rockefeller University Press</publisher><subject>Animals ; Cell Line - chemistry ; Drug Interactions ; Drug Synergism ; Epithelial Sodium Channels ; Immediate-Early Proteins ; Membrane Proteins - isolation & purification ; Membrane Proteins - pharmacology ; Membrane Proteins - physiology ; Mice ; Nuclear Proteins ; Oocytes ; Protein-Serine-Threonine Kinases - pharmacology ; Protein-Serine-Threonine Kinases - physiology ; Rats ; Serine Endopeptidases - isolation & purification ; Serine Endopeptidases - pharmacology ; Serine Endopeptidases - physiology ; Serpins ; Sodium Channels - drug effects ; Sodium Channels - metabolism ; Xenopus</subject><ispartof>The Journal of general physiology, 2002-08, Vol.120 (2), p.191-201</ispartof><rights>Copyright © 2002, The Rockefeller University Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c437t-58d7b2b674ae5513c1c482bfd74a4c5bcae59c93e369e9462b9f39548c2c7eae3</citedby><cites>FETCH-LOGICAL-c437t-58d7b2b674ae5513c1c482bfd74a4c5bcae59c93e369e9462b9f39548c2c7eae3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12149280$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Vuagniaux, Grégoire</creatorcontrib><creatorcontrib>Vallet, Véronique</creatorcontrib><creatorcontrib>Jaeger, Nicole Fowler</creatorcontrib><creatorcontrib>Hummler, Edith</creatorcontrib><creatorcontrib>Rossier, Bernard C</creatorcontrib><title>Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes</title><title>The Journal of general physiology</title><addtitle>J Gen Physiol</addtitle><description>Sodium balance is maintained by the precise regulation of the activity of the epithelial sodium channel (ENaC) in the kidney. We have recently reported an extracellular activation of ENaC-mediated sodium transport (I(Na)) by a GPI-anchored serine protease (mouse channel-activating protein, mCAP1) that was isolated from a cortical collecting duct cell line derived from mouse kidney. In the present study, we have identified two additional membrane-bound serine proteases (mCAP2 and mCAP3) that are expressed in the same cell line. We show that each of these proteases is able to increase I(Na) 6-10-fold in the Xenopus oocyte expression system. I(Na) and the number (N) of channels expressed at the cell surface (measured by binding of a FLAG monoclonal I(125)-radioiodinated antibody) were measured in the same oocyte. Using this assay, we show that mCAP1 increases I(Na) 10-fold (P < 0.001) but N remained unchanged (P = 0.9), indicating that mCAP1 regulates ENaC activity by increasing its average open probability of the whole cell (wcP(o)). The serum- and glucocorticoid-regulated kinase (Sgk1) involved in the aldosterone-dependent signaling cascade enhances I(Na) by 2.5-fold (P < 0.001) and N by 1.6-fold (P < 0.001), indicating a dual effect on N and wcP(o). Compared with Sgk1 alone, coexpression of Sgk1 with mCAP1 leads to a ninefold increase in I(Na) (P < 0.001) and 1.3-fold in N (P < 0.02). Similar results were observed for mCAP2 and mCAP3. The synergism between CAPs and Sgk1 on I(Na) was always more than additive, indicating a true potentiation. The synergistic effect of the two activation pathways allows a large dynamic range for ENaC-mediated sodium regulation crucial for a tight control of sodium homeostasis.</description><subject>Animals</subject><subject>Cell Line - chemistry</subject><subject>Drug Interactions</subject><subject>Drug Synergism</subject><subject>Epithelial Sodium Channels</subject><subject>Immediate-Early Proteins</subject><subject>Membrane Proteins - isolation & purification</subject><subject>Membrane Proteins - pharmacology</subject><subject>Membrane Proteins - physiology</subject><subject>Mice</subject><subject>Nuclear Proteins</subject><subject>Oocytes</subject><subject>Protein-Serine-Threonine Kinases - pharmacology</subject><subject>Protein-Serine-Threonine Kinases - physiology</subject><subject>Rats</subject><subject>Serine Endopeptidases - isolation & purification</subject><subject>Serine Endopeptidases - pharmacology</subject><subject>Serine Endopeptidases - physiology</subject><subject>Serpins</subject><subject>Sodium Channels - drug effects</subject><subject>Sodium Channels - metabolism</subject><subject>Xenopus</subject><issn>0022-1295</issn><issn>1540-7748</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVUctq3DAUFaWlmabddV20TGCU6jm2NoUwpA8ITSEtdGdk-dqjxJaMJAfm4_pv1eTRx93cw7nnHAkOQm8ZPWO0Vu9vhvmMU8prpetnaMWUpKSqZP0crQrLCeNaHaFXKd3QMorTl-iIcSY1r-kK_bree4iDS9lZbGx2dya74HHo8cVXs8XtHuddBMATTG00HkgbFt9huzPew0ieLH7ACaLzgOcYMpgECZ9M2_NvbI0Pi6-xKbYDFKf3sMiXidzDYVxssCGWPwTXkQjDMpoMHb51viThk-vhlp1i5_FP8GFeEr4Kdp8hvUYvejMmePO4j9GPjxfft5_J5dWnL9vzS2KlqDJRdVe1vN1U0oBSTFhmZc3bviuEtKq1hdZWCxAbDVpueKt7oZWsLbcVGBDH6MND7ry0E3QWfI5mbOboJhP3TTCu-f_i3a4Zwl3DuZBSVSVg_RBgY0gpQv_Hy2hzqLEpNTZPNRb5u3_f-yt-7E38BlKznEg</recordid><startdate>20020801</startdate><enddate>20020801</enddate><creator>Vuagniaux, Grégoire</creator><creator>Vallet, Véronique</creator><creator>Jaeger, Nicole Fowler</creator><creator>Hummler, Edith</creator><creator>Rossier, Bernard C</creator><general>The Rockefeller University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>5PM</scope></search><sort><creationdate>20020801</creationdate><title>Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes</title><author>Vuagniaux, Grégoire ; Vallet, Véronique ; Jaeger, Nicole Fowler ; Hummler, Edith ; Rossier, Bernard C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c437t-58d7b2b674ae5513c1c482bfd74a4c5bcae59c93e369e9462b9f39548c2c7eae3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Animals</topic><topic>Cell Line - chemistry</topic><topic>Drug Interactions</topic><topic>Drug Synergism</topic><topic>Epithelial Sodium Channels</topic><topic>Immediate-Early Proteins</topic><topic>Membrane Proteins - isolation & purification</topic><topic>Membrane Proteins - pharmacology</topic><topic>Membrane Proteins - physiology</topic><topic>Mice</topic><topic>Nuclear Proteins</topic><topic>Oocytes</topic><topic>Protein-Serine-Threonine Kinases - pharmacology</topic><topic>Protein-Serine-Threonine Kinases - physiology</topic><topic>Rats</topic><topic>Serine Endopeptidases - isolation & purification</topic><topic>Serine Endopeptidases - pharmacology</topic><topic>Serine Endopeptidases - physiology</topic><topic>Serpins</topic><topic>Sodium Channels - drug effects</topic><topic>Sodium Channels - metabolism</topic><topic>Xenopus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Vuagniaux, Grégoire</creatorcontrib><creatorcontrib>Vallet, Véronique</creatorcontrib><creatorcontrib>Jaeger, Nicole Fowler</creatorcontrib><creatorcontrib>Hummler, Edith</creatorcontrib><creatorcontrib>Rossier, Bernard C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of general physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Vuagniaux, Grégoire</au><au>Vallet, Véronique</au><au>Jaeger, Nicole Fowler</au><au>Hummler, Edith</au><au>Rossier, Bernard C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes</atitle><jtitle>The Journal of general physiology</jtitle><addtitle>J Gen Physiol</addtitle><date>2002-08-01</date><risdate>2002</risdate><volume>120</volume><issue>2</issue><spage>191</spage><epage>201</epage><pages>191-201</pages><issn>0022-1295</issn><eissn>1540-7748</eissn><abstract>Sodium balance is maintained by the precise regulation of the activity of the epithelial sodium channel (ENaC) in the kidney. We have recently reported an extracellular activation of ENaC-mediated sodium transport (I(Na)) by a GPI-anchored serine protease (mouse channel-activating protein, mCAP1) that was isolated from a cortical collecting duct cell line derived from mouse kidney. In the present study, we have identified two additional membrane-bound serine proteases (mCAP2 and mCAP3) that are expressed in the same cell line. We show that each of these proteases is able to increase I(Na) 6-10-fold in the Xenopus oocyte expression system. I(Na) and the number (N) of channels expressed at the cell surface (measured by binding of a FLAG monoclonal I(125)-radioiodinated antibody) were measured in the same oocyte. Using this assay, we show that mCAP1 increases I(Na) 10-fold (P < 0.001) but N remained unchanged (P = 0.9), indicating that mCAP1 regulates ENaC activity by increasing its average open probability of the whole cell (wcP(o)). The serum- and glucocorticoid-regulated kinase (Sgk1) involved in the aldosterone-dependent signaling cascade enhances I(Na) by 2.5-fold (P < 0.001) and N by 1.6-fold (P < 0.001), indicating a dual effect on N and wcP(o). Compared with Sgk1 alone, coexpression of Sgk1 with mCAP1 leads to a ninefold increase in I(Na) (P < 0.001) and 1.3-fold in N (P < 0.02). Similar results were observed for mCAP2 and mCAP3. The synergism between CAPs and Sgk1 on I(Na) was always more than additive, indicating a true potentiation. The synergistic effect of the two activation pathways allows a large dynamic range for ENaC-mediated sodium regulation crucial for a tight control of sodium homeostasis.</abstract><cop>United States</cop><pub>The Rockefeller University Press</pub><pmid>12149280</pmid><doi>10.1085/jgp.20028598</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Cell Line - chemistry Drug Interactions Drug Synergism Epithelial Sodium Channels Immediate-Early Proteins Membrane Proteins - isolation & purification Membrane Proteins - pharmacology Membrane Proteins - physiology Mice Nuclear Proteins Oocytes Protein-Serine-Threonine Kinases - pharmacology Protein-Serine-Threonine Kinases - physiology Rats Serine Endopeptidases - isolation & purification Serine Endopeptidases - pharmacology Serine Endopeptidases - physiology Serpins Sodium Channels - drug effects Sodium Channels - metabolism Xenopus |
title | Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes |
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