Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein
The Escherichia coli gene kch, similar in primary structure to eukaryotic voltage-gated potassium channels, was cloned and overexpressed in E. coli. The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or N-laurylsarcosine and was purified in mi...
Gespeichert in:
| Veröffentlicht in: | FEBS letters 1998-06, Vol.429 (1), p.104-108 |
|---|---|
| Hauptverfasser: | , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 108 |
|---|---|
| container_issue | 1 |
| container_start_page | 104 |
| container_title | FEBS letters |
| container_volume | 429 |
| creator | Voges, Dieter Jap, Bing K |
| description | The
Escherichia coli gene
kch, similar in primary structure to eukaryotic voltage-gated potassium channels, was cloned and overexpressed in
E. coli. The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or
N-laurylsarcosine and was purified in milligram amounts by imidazole elution from a nickel-chelate column. The molecular mass of the purified protein in a number of detergents with 12 carbon atom chains suggests that rKch forms primarily tetramers of the 50 kDa monomers. CD spectroscopy of the purified protein indicates a significant α-helical content that is preserved upon addition of SDS. |
| doi_str_mv | 10.1016/S0014-5793(98)00509-2 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_79987486</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0014579398005092</els_id><sourcerecordid>79987486</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4042-73de990f7db828d231524b1f7271f74889d96cc1a47886430e3030bc66e30c5e3</originalsourceid><addsrcrecordid>eNqNUF1r1TAYDqLMs82fMMiVKKwuH22TXImOMycbCE6vQ5q-5UTapCbt5vbrTU8Pu9WbJO_z9YYHoTNKPlBC64s7QmhZVELxd0q-J6QiqmAv0IZKwQte1vIl2jxLXqPjlH6RPEuqjtCRqivBFdugh-9gw9A4b_yE4c8YISUX_Dke5-g6Z82UJ2x8i-3ORGMniO5pBUOHb-zuHJusnTJ0D3ib7C4L7M4ZbEPv8BgmkwPnYbF7Dz0eY5jA-VP0qjN9gjeH-wT9vNr-uLwubr99-Xr56bawJSlZIXgLSpFOtI1ksmWcVqxsaCeYyEcppWpVbS01pZCyLjkBTjhpbF3nh62An6C3a27e-3uGNOnBJQt9bzyEOWmhlMw5dRZWq9DGkFKETo_RDSY-akr0UrjeF66XNrWSel-4Ztl3dlgwNwO0z65Dw5m_XvkH18Pj_4Xqq-1ntmcWQsk9vER9XKMgF3bvIOpkHXgLrYtgJ90G94_P_gXmUaVM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>79987486</pqid></control><display><type>article</type><title>Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein</title><source>Wiley Free Content</source><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><source>Elsevier ScienceDirect Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Voges, Dieter ; Jap, Bing K</creator><creatorcontrib>Voges, Dieter ; Jap, Bing K</creatorcontrib><description>The
Escherichia coli gene
kch, similar in primary structure to eukaryotic voltage-gated potassium channels, was cloned and overexpressed in
E. coli. The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or
N-laurylsarcosine and was purified in milligram amounts by imidazole elution from a nickel-chelate column. The molecular mass of the purified protein in a number of detergents with 12 carbon atom chains suggests that rKch forms primarily tetramers of the 50 kDa monomers. CD spectroscopy of the purified protein indicates a significant α-helical content that is preserved upon addition of SDS.</description><identifier>ISSN: 0014-5793</identifier><identifier>EISSN: 1873-3468</identifier><identifier>DOI: 10.1016/S0014-5793(98)00509-2</identifier><identifier>PMID: 9657392</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Bacterial Proteins - chemistry ; Bacterial Proteins - genetics ; Bacterial Proteins - isolation & purification ; Channel protein ; DDM, dodecyl-β-d-maltopyranoside ; Escherichia coli - chemistry ; Escherichia coli - genetics ; Escherichia coli Proteins ; IPTG, isopropyl-β-d-thiogalactopyranoside ; Kch ; LDAO, laurydimethylamine oxide ; Membrane protein ; Membrane Proteins - isolation & purification ; PCR, polymerase chain reaction ; Potassium channel ; Potassium Channels - chemistry ; Potassium Channels - genetics ; Potassium Channels - isolation & purification ; rKch, recombinant Kch ; sXXX kDa, ucXXX kDa, apparent molecular weight according to molecular sieve chromatography and analytical ultracentrifugation, respectively</subject><ispartof>FEBS letters, 1998-06, Vol.429 (1), p.104-108</ispartof><rights>1998 Federation of European Biochemical Societies</rights><rights>FEBS Letters 429 (1998) 1873-3468 © 2015 Federation of European Biochemical Societies</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4042-73de990f7db828d231524b1f7271f74889d96cc1a47886430e3030bc66e30c5e3</citedby><cites>FETCH-LOGICAL-c4042-73de990f7db828d231524b1f7271f74889d96cc1a47886430e3030bc66e30c5e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1016%2FS0014-5793%2898%2900509-2$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0014579398005092$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,1411,1427,3537,27901,27902,45550,45551,46384,46808,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9657392$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Voges, Dieter</creatorcontrib><creatorcontrib>Jap, Bing K</creatorcontrib><title>Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein</title><title>FEBS letters</title><addtitle>FEBS Lett</addtitle><description>The
Escherichia coli gene
kch, similar in primary structure to eukaryotic voltage-gated potassium channels, was cloned and overexpressed in
E. coli. The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or
N-laurylsarcosine and was purified in milligram amounts by imidazole elution from a nickel-chelate column. The molecular mass of the purified protein in a number of detergents with 12 carbon atom chains suggests that rKch forms primarily tetramers of the 50 kDa monomers. CD spectroscopy of the purified protein indicates a significant α-helical content that is preserved upon addition of SDS.</description><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - isolation & purification</subject><subject>Channel protein</subject><subject>DDM, dodecyl-β-d-maltopyranoside</subject><subject>Escherichia coli - chemistry</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli Proteins</subject><subject>IPTG, isopropyl-β-d-thiogalactopyranoside</subject><subject>Kch</subject><subject>LDAO, laurydimethylamine oxide</subject><subject>Membrane protein</subject><subject>Membrane Proteins - isolation & purification</subject><subject>PCR, polymerase chain reaction</subject><subject>Potassium channel</subject><subject>Potassium Channels - chemistry</subject><subject>Potassium Channels - genetics</subject><subject>Potassium Channels - isolation & purification</subject><subject>rKch, recombinant Kch</subject><subject>sXXX kDa, ucXXX kDa, apparent molecular weight according to molecular sieve chromatography and analytical ultracentrifugation, respectively</subject><issn>0014-5793</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNUF1r1TAYDqLMs82fMMiVKKwuH22TXImOMycbCE6vQ5q-5UTapCbt5vbrTU8Pu9WbJO_z9YYHoTNKPlBC64s7QmhZVELxd0q-J6QiqmAv0IZKwQte1vIl2jxLXqPjlH6RPEuqjtCRqivBFdugh-9gw9A4b_yE4c8YISUX_Dke5-g6Z82UJ2x8i-3ORGMniO5pBUOHb-zuHJusnTJ0D3ib7C4L7M4ZbEPv8BgmkwPnYbF7Dz0eY5jA-VP0qjN9gjeH-wT9vNr-uLwubr99-Xr56bawJSlZIXgLSpFOtI1ksmWcVqxsaCeYyEcppWpVbS01pZCyLjkBTjhpbF3nh62An6C3a27e-3uGNOnBJQt9bzyEOWmhlMw5dRZWq9DGkFKETo_RDSY-akr0UrjeF66XNrWSel-4Ztl3dlgwNwO0z65Dw5m_XvkH18Pj_4Xqq-1ntmcWQsk9vER9XKMgF3bvIOpkHXgLrYtgJ90G94_P_gXmUaVM</recordid><startdate>19980605</startdate><enddate>19980605</enddate><creator>Voges, Dieter</creator><creator>Jap, Bing K</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</scope><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>19980605</creationdate><title>Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein</title><author>Voges, Dieter ; Jap, Bing K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4042-73de990f7db828d231524b1f7271f74889d96cc1a47886430e3030bc66e30c5e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - isolation & purification</topic><topic>Channel protein</topic><topic>DDM, dodecyl-β-d-maltopyranoside</topic><topic>Escherichia coli - chemistry</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli Proteins</topic><topic>IPTG, isopropyl-β-d-thiogalactopyranoside</topic><topic>Kch</topic><topic>LDAO, laurydimethylamine oxide</topic><topic>Membrane protein</topic><topic>Membrane Proteins - isolation & purification</topic><topic>PCR, polymerase chain reaction</topic><topic>Potassium channel</topic><topic>Potassium Channels - chemistry</topic><topic>Potassium Channels - genetics</topic><topic>Potassium Channels - isolation & purification</topic><topic>rKch, recombinant Kch</topic><topic>sXXX kDa, ucXXX kDa, apparent molecular weight according to molecular sieve chromatography and analytical ultracentrifugation, respectively</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Voges, Dieter</creatorcontrib><creatorcontrib>Jap, Bing K</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><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>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Voges, Dieter</au><au>Jap, Bing K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>1998-06-05</date><risdate>1998</risdate><volume>429</volume><issue>1</issue><spage>104</spage><epage>108</epage><pages>104-108</pages><issn>0014-5793</issn><eissn>1873-3468</eissn><abstract>The
Escherichia coli gene
kch, similar in primary structure to eukaryotic voltage-gated potassium channels, was cloned and overexpressed in
E. coli. The protein was solubilized from the plasma membrane with dodecylmaltopyranoside, lauryldimethylamine oxide or
N-laurylsarcosine and was purified in milligram amounts by imidazole elution from a nickel-chelate column. The molecular mass of the purified protein in a number of detergents with 12 carbon atom chains suggests that rKch forms primarily tetramers of the 50 kDa monomers. CD spectroscopy of the purified protein indicates a significant α-helical content that is preserved upon addition of SDS.</abstract><cop>England</cop><pub>Elsevier B.V</pub><pmid>9657392</pmid><doi>10.1016/S0014-5793(98)00509-2</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0014-5793 |
| ispartof | FEBS letters, 1998-06, Vol.429 (1), p.104-108 |
| issn | 0014-5793 1873-3468 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79987486 |
| source | Wiley Free Content; MEDLINE; Wiley Online Library Journals Frontfile Complete; Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - isolation & purification Channel protein DDM, dodecyl-β-d-maltopyranoside Escherichia coli - chemistry Escherichia coli - genetics Escherichia coli Proteins IPTG, isopropyl-β-d-thiogalactopyranoside Kch LDAO, laurydimethylamine oxide Membrane protein Membrane Proteins - isolation & purification PCR, polymerase chain reaction Potassium channel Potassium Channels - chemistry Potassium Channels - genetics Potassium Channels - isolation & purification rKch, recombinant Kch sXXX kDa, ucXXX kDa, apparent molecular weight according to molecular sieve chromatography and analytical ultracentrifugation, respectively |
| title | Recombinant expression, purification and characterization of Kch, a putative Escherichia coli potassium channel protein |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-08T21%3A47%3A39IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Recombinant%20expression,%20purification%20and%20characterization%20of%20Kch,%20a%20putative%20Escherichia%20coli%20potassium%20channel%20protein&rft.jtitle=FEBS%20letters&rft.au=Voges,%20Dieter&rft.date=1998-06-05&rft.volume=429&rft.issue=1&rft.spage=104&rft.epage=108&rft.pages=104-108&rft.issn=0014-5793&rft.eissn=1873-3468&rft_id=info:doi/10.1016/S0014-5793(98)00509-2&rft_dat=%3Cproquest_cross%3E79987486%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=79987486&rft_id=info:pmid/9657392&rft_els_id=S0014579398005092&rfr_iscdi=true |