IRBIT plays an important role in NHE3-mediated pHi regulation in HSG cells
•IRBIT plays a putative role in pHi regulation in HSG cells.•In addition to NHE1, NHE3 is also involved in pHi recovery in HSG cells.•IRBIT regulates pHi via its effect on membrane NHE3 translocation.•IRBIT-induced membrane NHE3 translocation is further regulated by SPAK. Expression of inositol-1,4,...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2013-07, Vol.437 (1), p.18-22 |
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| container_title | Biochemical and biophysical research communications |
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| creator | Tran, Tien Manh Park, Moon-Yong Lee, Jiyeon Bae, Jun-Seok Hwang, Sung-Min Choi, Se-Young Mikoshiba, Katsuhiko Park, Kyungpyo |
| description | •IRBIT plays a putative role in pHi regulation in HSG cells.•In addition to NHE1, NHE3 is also involved in pHi recovery in HSG cells.•IRBIT regulates pHi via its effect on membrane NHE3 translocation.•IRBIT-induced membrane NHE3 translocation is further regulated by SPAK.
Expression of inositol-1,4,5-trisphosphate (IP3) receptor-binding protein (IRBIT) has been reported in epithelial cells. However, its role in pHi regulation is not well understood. In this study, we investigated the role of IRBIT in pHi regulation, mediated by Na+/H+ exchangers (NHEs), in salivary glands. We measured pHi recovery from cell acidification in BCECF-loaded salivary HSG cells. Western blot and co-immunoprecipitation (CO-IP) assays were also performed, showing that NHE1, 2 and 3 are expressed, and IRBIT binds to NHE3. HOE642, a specific NHE1 blocker, inhibited pHi recovery, but 40% pHi recovery was still observed even at the highest concentration of HOE642. Furthermore, pretreatment of the cells with siIRBIT significantly inhibited pHi recovery, indicating that NHE3 potentially plays a role in pHi recovery as well. The amount of membrane-localized NHE3 and its interaction with IRBIT are also significantly increased by cell acidification. In addition, we found that Ste20p-related proline alanine-rich kinase (SPAK) reverses the effect of IRBIT on membrane NHE3 translocation. Taken together, we conclude that IRBIT plays an important role in pHi regulation, mediated by NHE3, and further regulated by SPAK. |
| doi_str_mv | 10.1016/j.bbrc.2013.06.010 |
| format | Article |
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Expression of inositol-1,4,5-trisphosphate (IP3) receptor-binding protein (IRBIT) has been reported in epithelial cells. However, its role in pHi regulation is not well understood. In this study, we investigated the role of IRBIT in pHi regulation, mediated by Na+/H+ exchangers (NHEs), in salivary glands. We measured pHi recovery from cell acidification in BCECF-loaded salivary HSG cells. Western blot and co-immunoprecipitation (CO-IP) assays were also performed, showing that NHE1, 2 and 3 are expressed, and IRBIT binds to NHE3. HOE642, a specific NHE1 blocker, inhibited pHi recovery, but 40% pHi recovery was still observed even at the highest concentration of HOE642. Furthermore, pretreatment of the cells with siIRBIT significantly inhibited pHi recovery, indicating that NHE3 potentially plays a role in pHi recovery as well. The amount of membrane-localized NHE3 and its interaction with IRBIT are also significantly increased by cell acidification. In addition, we found that Ste20p-related proline alanine-rich kinase (SPAK) reverses the effect of IRBIT on membrane NHE3 translocation. Taken together, we conclude that IRBIT plays an important role in pHi regulation, mediated by NHE3, and further regulated by SPAK.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2013.06.010</identifier><identifier>PMID: 23769829</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Cation Transport Proteins - metabolism ; Cell Line ; HOE642 ; Humans ; Hydrogen-Ion Concentration ; Intracellular Space - metabolism ; IRBIT ; Lectins, C-Type - metabolism ; Membrane Proteins - metabolism ; Na+/H+ exchangers ; pHi Recovery ; Protein Binding ; Protein Transport ; Protein-Serine-Threonine Kinases - metabolism ; Salivary Glands - cytology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchanger 3 ; Sodium-Hydrogen Exchangers - metabolism ; SPAK</subject><ispartof>Biochemical and biophysical research communications, 2013-07, Vol.437 (1), p.18-22</ispartof><rights>2013 Elsevier Inc.</rights><rights>Copyright © 2013 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c337t-566fb088d6ae365526638b8c4d22a3c6cee2f3659042e3fe5fa1ef8f3e3b89fd3</citedby><cites>FETCH-LOGICAL-c337t-566fb088d6ae365526638b8c4d22a3c6cee2f3659042e3fe5fa1ef8f3e3b89fd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2013.06.010$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23769829$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tran, Tien Manh</creatorcontrib><creatorcontrib>Park, Moon-Yong</creatorcontrib><creatorcontrib>Lee, Jiyeon</creatorcontrib><creatorcontrib>Bae, Jun-Seok</creatorcontrib><creatorcontrib>Hwang, Sung-Min</creatorcontrib><creatorcontrib>Choi, Se-Young</creatorcontrib><creatorcontrib>Mikoshiba, Katsuhiko</creatorcontrib><creatorcontrib>Park, Kyungpyo</creatorcontrib><title>IRBIT plays an important role in NHE3-mediated pHi regulation in HSG cells</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>•IRBIT plays a putative role in pHi regulation in HSG cells.•In addition to NHE1, NHE3 is also involved in pHi recovery in HSG cells.•IRBIT regulates pHi via its effect on membrane NHE3 translocation.•IRBIT-induced membrane NHE3 translocation is further regulated by SPAK.
Expression of inositol-1,4,5-trisphosphate (IP3) receptor-binding protein (IRBIT) has been reported in epithelial cells. However, its role in pHi regulation is not well understood. In this study, we investigated the role of IRBIT in pHi regulation, mediated by Na+/H+ exchangers (NHEs), in salivary glands. We measured pHi recovery from cell acidification in BCECF-loaded salivary HSG cells. Western blot and co-immunoprecipitation (CO-IP) assays were also performed, showing that NHE1, 2 and 3 are expressed, and IRBIT binds to NHE3. HOE642, a specific NHE1 blocker, inhibited pHi recovery, but 40% pHi recovery was still observed even at the highest concentration of HOE642. Furthermore, pretreatment of the cells with siIRBIT significantly inhibited pHi recovery, indicating that NHE3 potentially plays a role in pHi recovery as well. The amount of membrane-localized NHE3 and its interaction with IRBIT are also significantly increased by cell acidification. In addition, we found that Ste20p-related proline alanine-rich kinase (SPAK) reverses the effect of IRBIT on membrane NHE3 translocation. Taken together, we conclude that IRBIT plays an important role in pHi regulation, mediated by NHE3, and further regulated by SPAK.</description><subject>Cation Transport Proteins - metabolism</subject><subject>Cell Line</subject><subject>HOE642</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Intracellular Space - metabolism</subject><subject>IRBIT</subject><subject>Lectins, C-Type - metabolism</subject><subject>Membrane Proteins - metabolism</subject><subject>Na+/H+ exchangers</subject><subject>pHi Recovery</subject><subject>Protein Binding</subject><subject>Protein Transport</subject><subject>Protein-Serine-Threonine Kinases - metabolism</subject><subject>Salivary Glands - cytology</subject><subject>Sodium-Hydrogen Exchanger 1</subject><subject>Sodium-Hydrogen Exchanger 3</subject><subject>Sodium-Hydrogen Exchangers - metabolism</subject><subject>SPAK</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9LwzAYxoMoOqdfwIPk6KX1TdJlLXjRMbfJUNAJ3kKavpGMrq1JJ-zb27Lp0dN7eP7wPj9CrhjEDJi8Xcd57k3MgYkYZAwMjsiAQQYRZ5AckwEAyIhn7OOMnIewBmAskdkpOeNiLLOUZwPytHh9WKxoU-pdoLqibtPUvtVVS31dInUVfZ5PRbTBwukWC9rMHfX4uS116-qq1-dvM2qwLMMFObG6DHh5uEPy_jhdTebR8mW2mNwvIyPEuI1GUtoc0rSQGoUcjbiUIs1TkxSca2GkQeS2EzJIOAqLI6sZ2tQKFHma2UIMyc2-t_H11xZDqzYu9B_oCuttUCxhTHKZMtFZ-d5qfB2CR6sa7zba7xQD1TNUa9UzVD1DBVJ1DLvQ9aF_m3e7_yK_0DrD3d6A3cpvh14F47AyHSOPplVF7f7r_wGrioDW</recordid><startdate>20130719</startdate><enddate>20130719</enddate><creator>Tran, Tien Manh</creator><creator>Park, Moon-Yong</creator><creator>Lee, Jiyeon</creator><creator>Bae, Jun-Seok</creator><creator>Hwang, Sung-Min</creator><creator>Choi, Se-Young</creator><creator>Mikoshiba, Katsuhiko</creator><creator>Park, Kyungpyo</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>20130719</creationdate><title>IRBIT plays an important role in NHE3-mediated pHi regulation in HSG cells</title><author>Tran, Tien Manh ; Park, Moon-Yong ; Lee, Jiyeon ; Bae, Jun-Seok ; Hwang, Sung-Min ; Choi, Se-Young ; Mikoshiba, Katsuhiko ; Park, Kyungpyo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c337t-566fb088d6ae365526638b8c4d22a3c6cee2f3659042e3fe5fa1ef8f3e3b89fd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Cation Transport Proteins - metabolism</topic><topic>Cell Line</topic><topic>HOE642</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Intracellular Space - metabolism</topic><topic>IRBIT</topic><topic>Lectins, C-Type - metabolism</topic><topic>Membrane Proteins - metabolism</topic><topic>Na+/H+ exchangers</topic><topic>pHi Recovery</topic><topic>Protein Binding</topic><topic>Protein Transport</topic><topic>Protein-Serine-Threonine Kinases - metabolism</topic><topic>Salivary Glands - cytology</topic><topic>Sodium-Hydrogen Exchanger 1</topic><topic>Sodium-Hydrogen Exchanger 3</topic><topic>Sodium-Hydrogen Exchangers - metabolism</topic><topic>SPAK</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tran, Tien Manh</creatorcontrib><creatorcontrib>Park, Moon-Yong</creatorcontrib><creatorcontrib>Lee, Jiyeon</creatorcontrib><creatorcontrib>Bae, Jun-Seok</creatorcontrib><creatorcontrib>Hwang, Sung-Min</creatorcontrib><creatorcontrib>Choi, Se-Young</creatorcontrib><creatorcontrib>Mikoshiba, Katsuhiko</creatorcontrib><creatorcontrib>Park, Kyungpyo</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tran, Tien Manh</au><au>Park, Moon-Yong</au><au>Lee, Jiyeon</au><au>Bae, Jun-Seok</au><au>Hwang, Sung-Min</au><au>Choi, Se-Young</au><au>Mikoshiba, Katsuhiko</au><au>Park, Kyungpyo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>IRBIT plays an important role in NHE3-mediated pHi regulation in HSG cells</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2013-07-19</date><risdate>2013</risdate><volume>437</volume><issue>1</issue><spage>18</spage><epage>22</epage><pages>18-22</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>•IRBIT plays a putative role in pHi regulation in HSG cells.•In addition to NHE1, NHE3 is also involved in pHi recovery in HSG cells.•IRBIT regulates pHi via its effect on membrane NHE3 translocation.•IRBIT-induced membrane NHE3 translocation is further regulated by SPAK.
Expression of inositol-1,4,5-trisphosphate (IP3) receptor-binding protein (IRBIT) has been reported in epithelial cells. However, its role in pHi regulation is not well understood. In this study, we investigated the role of IRBIT in pHi regulation, mediated by Na+/H+ exchangers (NHEs), in salivary glands. We measured pHi recovery from cell acidification in BCECF-loaded salivary HSG cells. Western blot and co-immunoprecipitation (CO-IP) assays were also performed, showing that NHE1, 2 and 3 are expressed, and IRBIT binds to NHE3. HOE642, a specific NHE1 blocker, inhibited pHi recovery, but 40% pHi recovery was still observed even at the highest concentration of HOE642. Furthermore, pretreatment of the cells with siIRBIT significantly inhibited pHi recovery, indicating that NHE3 potentially plays a role in pHi recovery as well. The amount of membrane-localized NHE3 and its interaction with IRBIT are also significantly increased by cell acidification. In addition, we found that Ste20p-related proline alanine-rich kinase (SPAK) reverses the effect of IRBIT on membrane NHE3 translocation. Taken together, we conclude that IRBIT plays an important role in pHi regulation, mediated by NHE3, and further regulated by SPAK.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23769829</pmid><doi>10.1016/j.bbrc.2013.06.010</doi><tpages>5</tpages></addata></record> |
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| ispartof | Biochemical and biophysical research communications, 2013-07, Vol.437 (1), p.18-22 |
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| language | eng |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Cation Transport Proteins - metabolism Cell Line HOE642 Humans Hydrogen-Ion Concentration Intracellular Space - metabolism IRBIT Lectins, C-Type - metabolism Membrane Proteins - metabolism Na+/H+ exchangers pHi Recovery Protein Binding Protein Transport Protein-Serine-Threonine Kinases - metabolism Salivary Glands - cytology Sodium-Hydrogen Exchanger 1 Sodium-Hydrogen Exchanger 3 Sodium-Hydrogen Exchangers - metabolism SPAK |
| title | IRBIT plays an important role in NHE3-mediated pHi regulation in HSG cells |
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