Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells
Cysteine string protein-α (CSPα) is a chaperone to ensure protein folding. Loss of CSPα function associates with many neurological diseases. However, its function in modulating regulated exocytosis remains elusive. Although cspα-knockouts exhibit impaired synaptic transmission, overexpression of CSP...
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| description | Cysteine string protein-α (CSPα) is a chaperone to ensure protein folding. Loss of CSPα function associates with many neurological diseases. However, its function in modulating regulated exocytosis remains elusive. Although cspα-knockouts exhibit impaired synaptic transmission, overexpression of CSPα in neuroendocrine cells inhibits secretion. These seemingly conflicting results lead to a hypothesis that CSPα may undergo a modification that switches its function in regulating neurotransmitter and hormone secretion. Previous studies implied that CSPα undergoes phosphorylation at Ser10 that may influence exocytosis by altering fusion pore dynamics. However, direct evidence is missing up to date.
Using amperometry, we investigated how phosphorylation at Ser10 of CSPα (CSPα-Ser10) modulates regulated exocytosis and if this modulation involves regulating a specific kinetic step of fusion pore dynamics. The real-time exocytosis of single vesicles was detected in PC12 cells overexpressing control vector, wild-type CSPα (WT), the CSPα phosphodeficient mutant (S10A), or the CSPα phosphomimetic mutants (S10D and S10E). The shapes of amperometric signals were used to distinguish the full-fusion events (i.e., prespike feet followed by spikes) and the kiss-and-run events (i.e., square-shaped flickers). We found that the secretion rate was significantly increased in cells overexpressing S10D or S10E compared to WT or S10A. Further analysis showed that overexpression of S10D or S10E prolonged fusion pore lifetime compared to WT or S10A. The fraction of kiss-and-run events was significantly lower but the frequency of full-fusion events was higher in cells overexpressing S10D or S10E compared to WT or S10A. Advanced kinetic analysis suggests that overexpression of S10D or S10E may stabilize open fusion pores mainly by inhibiting them from closing.
CSPα may modulate fusion pore dynamics in a phosphorylation-dependent manner. Therefore, through changing its phosphorylated state influenced by diverse cellular signalings, CSPα may have a great capacity to modulate the rate of regulated exocytosis. |
| doi_str_mv | 10.1371/journal.pone.0099180 |
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Using amperometry, we investigated how phosphorylation at Ser10 of CSPα (CSPα-Ser10) modulates regulated exocytosis and if this modulation involves regulating a specific kinetic step of fusion pore dynamics. The real-time exocytosis of single vesicles was detected in PC12 cells overexpressing control vector, wild-type CSPα (WT), the CSPα phosphodeficient mutant (S10A), or the CSPα phosphomimetic mutants (S10D and S10E). The shapes of amperometric signals were used to distinguish the full-fusion events (i.e., prespike feet followed by spikes) and the kiss-and-run events (i.e., square-shaped flickers). We found that the secretion rate was significantly increased in cells overexpressing S10D or S10E compared to WT or S10A. Further analysis showed that overexpression of S10D or S10E prolonged fusion pore lifetime compared to WT or S10A. The fraction of kiss-and-run events was significantly lower but the frequency of full-fusion events was higher in cells overexpressing S10D or S10E compared to WT or S10A. Advanced kinetic analysis suggests that overexpression of S10D or S10E may stabilize open fusion pores mainly by inhibiting them from closing.
CSPα may modulate fusion pore dynamics in a phosphorylation-dependent manner. Therefore, through changing its phosphorylated state influenced by diverse cellular signalings, CSPα may have a great capacity to modulate the rate of regulated exocytosis.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0099180</identifier><identifier>PMID: 24956274</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amperometry ; Animals ; Biology and Life Sciences ; Cell Membrane - metabolism ; Cellular biology ; Cysteine ; Disease transmission ; Drosophila ; Dynamics ; Electrical measurement ; Endocytosis ; Exocytosis ; HSP40 Heat-Shock Proteins - genetics ; HSP40 Heat-Shock Proteins - secretion ; Insects ; Kinases ; Kinetics ; Life sciences ; Mammals ; Membrane Fusion ; Membrane Proteins - genetics ; Membrane Proteins - secretion ; Mutant Proteins - secretion ; Mutants ; Mutation ; Mutation - genetics ; Neurobiology ; Neurodegeneration ; Neurological diseases ; Neurosciences ; PC12 Cells ; Pheochromocytoma cells ; Phosphorylation ; Physiology ; Protein folding ; Proteins ; Rats ; Research and Analysis Methods ; Rodents ; Secretion ; String protein ; Switches ; Synaptic transmission</subject><ispartof>PloS one, 2014-06, Vol.9 (6), p.e99180-e99180</ispartof><rights>2014 Chiang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Chiang et al 2014 Chiang et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-491192db757a8c6dbcc9d501be2008b7aa331f712c7b136bebb4a9cc9b19a3893</citedby><cites>FETCH-LOGICAL-c526t-491192db757a8c6dbcc9d501be2008b7aa331f712c7b136bebb4a9cc9b19a3893</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067274/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067274/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,2096,2915,23847,27905,27906,53772,53774,79349,79350</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24956274$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Gasman, Stephane</contributor><creatorcontrib>Chiang, Ning</creatorcontrib><creatorcontrib>Hsiao, Yu-Tien</creatorcontrib><creatorcontrib>Yang, Hui-Ju</creatorcontrib><creatorcontrib>Lin, Yu-Chun</creatorcontrib><creatorcontrib>Lu, Juu-Chin</creatorcontrib><creatorcontrib>Wang, Chih-Tien</creatorcontrib><title>Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Cysteine string protein-α (CSPα) is a chaperone to ensure protein folding. Loss of CSPα function associates with many neurological diseases. However, its function in modulating regulated exocytosis remains elusive. Although cspα-knockouts exhibit impaired synaptic transmission, overexpression of CSPα in neuroendocrine cells inhibits secretion. These seemingly conflicting results lead to a hypothesis that CSPα may undergo a modification that switches its function in regulating neurotransmitter and hormone secretion. Previous studies implied that CSPα undergoes phosphorylation at Ser10 that may influence exocytosis by altering fusion pore dynamics. However, direct evidence is missing up to date.
Using amperometry, we investigated how phosphorylation at Ser10 of CSPα (CSPα-Ser10) modulates regulated exocytosis and if this modulation involves regulating a specific kinetic step of fusion pore dynamics. The real-time exocytosis of single vesicles was detected in PC12 cells overexpressing control vector, wild-type CSPα (WT), the CSPα phosphodeficient mutant (S10A), or the CSPα phosphomimetic mutants (S10D and S10E). The shapes of amperometric signals were used to distinguish the full-fusion events (i.e., prespike feet followed by spikes) and the kiss-and-run events (i.e., square-shaped flickers). We found that the secretion rate was significantly increased in cells overexpressing S10D or S10E compared to WT or S10A. Further analysis showed that overexpression of S10D or S10E prolonged fusion pore lifetime compared to WT or S10A. The fraction of kiss-and-run events was significantly lower but the frequency of full-fusion events was higher in cells overexpressing S10D or S10E compared to WT or S10A. Advanced kinetic analysis suggests that overexpression of S10D or S10E may stabilize open fusion pores mainly by inhibiting them from closing.
CSPα may modulate fusion pore dynamics in a phosphorylation-dependent manner. Therefore, through changing its phosphorylated state influenced by diverse cellular signalings, CSPα may have a great capacity to modulate the rate of regulated exocytosis.</description><subject>Amperometry</subject><subject>Animals</subject><subject>Biology and Life Sciences</subject><subject>Cell Membrane - metabolism</subject><subject>Cellular biology</subject><subject>Cysteine</subject><subject>Disease transmission</subject><subject>Drosophila</subject><subject>Dynamics</subject><subject>Electrical measurement</subject><subject>Endocytosis</subject><subject>Exocytosis</subject><subject>HSP40 Heat-Shock Proteins - genetics</subject><subject>HSP40 Heat-Shock Proteins - secretion</subject><subject>Insects</subject><subject>Kinases</subject><subject>Kinetics</subject><subject>Life sciences</subject><subject>Mammals</subject><subject>Membrane Fusion</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - secretion</subject><subject>Mutant Proteins - secretion</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Mutation - genetics</subject><subject>Neurobiology</subject><subject>Neurodegeneration</subject><subject>Neurological diseases</subject><subject>Neurosciences</subject><subject>PC12 Cells</subject><subject>Pheochromocytoma cells</subject><subject>Phosphorylation</subject><subject>Physiology</subject><subject>Protein folding</subject><subject>Proteins</subject><subject>Rats</subject><subject>Research and Analysis Methods</subject><subject>Rodents</subject><subject>Secretion</subject><subject>String protein</subject><subject>Switches</subject><subject>Synaptic transmission</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptUkuO1DAUjBCIGQZugMASGzZp_ImdeIOEWnxGGolZwNqynZdut5I42A6ij8FRuAhnwqEzoxnEyvZzVfm9chXFc4I3hNXkzcHPYdT9ZvIjbDCWkjT4QXFOJKOloJg9vLM_K57EeMCYs0aIx8UZrSQXtK7Oi5_Xex-nvR_cAMlZNMxJJ-dH5DtkjzGBGwHFFNy4Q1Pwy7n8_Qu50QbQESJKe0BBJ1gIAXZzn_ctgh_eHpOPLiJzRINvl_qi0c1xUZ98ANQeRz04G7Maut4Siiz0fXxaPOp0H-HZul4UXz-8_7L9VF59_ni5fXdVWk5FKitJiKStqXmtGytaY61sOSYGKMaNqbVmjHQ1obY2hAkDxlRaZpAhUrNGsovi5Ul36n1Uq5tREc5kJSuMRUZcnhCt1wc1BTfocFReO_W34MNO6ZBN6yGzLDfYCt7ypuo4GMsEz143YCnO7WWtt-trsxmgtTCmoPt7ovdvRrdXO_9dVVjU-aeywOtVIPhvM8SkBhcXw_QIfl76rjDBGUgz9NU_0P9PV51QNvgYA3S3zRCsloTdsNSSMLUmLNNe3B3klnQTKfYHOGLTGA</recordid><startdate>20140623</startdate><enddate>20140623</enddate><creator>Chiang, Ning</creator><creator>Hsiao, Yu-Tien</creator><creator>Yang, Hui-Ju</creator><creator>Lin, Yu-Chun</creator><creator>Lu, Juu-Chin</creator><creator>Wang, Chih-Tien</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20140623</creationdate><title>Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells</title><author>Chiang, Ning ; Hsiao, Yu-Tien ; Yang, Hui-Ju ; Lin, Yu-Chun ; Lu, Juu-Chin ; Wang, Chih-Tien</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-491192db757a8c6dbcc9d501be2008b7aa331f712c7b136bebb4a9cc9b19a3893</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Amperometry</topic><topic>Animals</topic><topic>Biology and Life Sciences</topic><topic>Cell Membrane - metabolism</topic><topic>Cellular biology</topic><topic>Cysteine</topic><topic>Disease transmission</topic><topic>Drosophila</topic><topic>Dynamics</topic><topic>Electrical measurement</topic><topic>Endocytosis</topic><topic>Exocytosis</topic><topic>HSP40 Heat-Shock Proteins - genetics</topic><topic>HSP40 Heat-Shock Proteins - secretion</topic><topic>Insects</topic><topic>Kinases</topic><topic>Kinetics</topic><topic>Life sciences</topic><topic>Mammals</topic><topic>Membrane Fusion</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - secretion</topic><topic>Mutant Proteins - secretion</topic><topic>Mutants</topic><topic>Mutation</topic><topic>Mutation - genetics</topic><topic>Neurobiology</topic><topic>Neurodegeneration</topic><topic>Neurological diseases</topic><topic>Neurosciences</topic><topic>PC12 Cells</topic><topic>Pheochromocytoma cells</topic><topic>Phosphorylation</topic><topic>Physiology</topic><topic>Protein folding</topic><topic>Proteins</topic><topic>Rats</topic><topic>Research and Analysis Methods</topic><topic>Rodents</topic><topic>Secretion</topic><topic>String protein</topic><topic>Switches</topic><topic>Synaptic transmission</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chiang, Ning</creatorcontrib><creatorcontrib>Hsiao, Yu-Tien</creatorcontrib><creatorcontrib>Yang, Hui-Ju</creatorcontrib><creatorcontrib>Lin, Yu-Chun</creatorcontrib><creatorcontrib>Lu, Juu-Chin</creatorcontrib><creatorcontrib>Wang, Chih-Tien</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chiang, Ning</au><au>Hsiao, Yu-Tien</au><au>Yang, Hui-Ju</au><au>Lin, Yu-Chun</au><au>Lu, Juu-Chin</au><au>Wang, Chih-Tien</au><au>Gasman, Stephane</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-06-23</date><risdate>2014</risdate><volume>9</volume><issue>6</issue><spage>e99180</spage><epage>e99180</epage><pages>e99180-e99180</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Cysteine string protein-α (CSPα) is a chaperone to ensure protein folding. Loss of CSPα function associates with many neurological diseases. However, its function in modulating regulated exocytosis remains elusive. Although cspα-knockouts exhibit impaired synaptic transmission, overexpression of CSPα in neuroendocrine cells inhibits secretion. These seemingly conflicting results lead to a hypothesis that CSPα may undergo a modification that switches its function in regulating neurotransmitter and hormone secretion. Previous studies implied that CSPα undergoes phosphorylation at Ser10 that may influence exocytosis by altering fusion pore dynamics. However, direct evidence is missing up to date.
Using amperometry, we investigated how phosphorylation at Ser10 of CSPα (CSPα-Ser10) modulates regulated exocytosis and if this modulation involves regulating a specific kinetic step of fusion pore dynamics. The real-time exocytosis of single vesicles was detected in PC12 cells overexpressing control vector, wild-type CSPα (WT), the CSPα phosphodeficient mutant (S10A), or the CSPα phosphomimetic mutants (S10D and S10E). The shapes of amperometric signals were used to distinguish the full-fusion events (i.e., prespike feet followed by spikes) and the kiss-and-run events (i.e., square-shaped flickers). We found that the secretion rate was significantly increased in cells overexpressing S10D or S10E compared to WT or S10A. Further analysis showed that overexpression of S10D or S10E prolonged fusion pore lifetime compared to WT or S10A. The fraction of kiss-and-run events was significantly lower but the frequency of full-fusion events was higher in cells overexpressing S10D or S10E compared to WT or S10A. Advanced kinetic analysis suggests that overexpression of S10D or S10E may stabilize open fusion pores mainly by inhibiting them from closing.
CSPα may modulate fusion pore dynamics in a phosphorylation-dependent manner. Therefore, through changing its phosphorylated state influenced by diverse cellular signalings, CSPα may have a great capacity to modulate the rate of regulated exocytosis.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24956274</pmid><doi>10.1371/journal.pone.0099180</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Amperometry Animals Biology and Life Sciences Cell Membrane - metabolism Cellular biology Cysteine Disease transmission Drosophila Dynamics Electrical measurement Endocytosis Exocytosis HSP40 Heat-Shock Proteins - genetics HSP40 Heat-Shock Proteins - secretion Insects Kinases Kinetics Life sciences Mammals Membrane Fusion Membrane Proteins - genetics Membrane Proteins - secretion Mutant Proteins - secretion Mutants Mutation Mutation - genetics Neurobiology Neurodegeneration Neurological diseases Neurosciences PC12 Cells Pheochromocytoma cells Phosphorylation Physiology Protein folding Proteins Rats Research and Analysis Methods Rodents Secretion String protein Switches Synaptic transmission |
| title | Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-18T18%3A52%3A21IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Phosphomimetic%20mutation%20of%20cysteine%20string%20protein-%CE%B1%20increases%20the%20rate%20of%20regulated%20exocytosis%20by%20modulating%20fusion%20pore%20dynamics%20in%20PC12%20cells&rft.jtitle=PloS%20one&rft.au=Chiang,%20Ning&rft.date=2014-06-23&rft.volume=9&rft.issue=6&rft.spage=e99180&rft.epage=e99180&rft.pages=e99180-e99180&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0099180&rft_dat=%3Cproquest_plos_%3E1540107432%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1539494006&rft_id=info:pmid/24956274&rft_doaj_id=oai_doaj_org_article_15c5b0c65d584f5ebc3658668ec208c6&rfr_iscdi=true |