Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies
The EGFR/HER receptor family of an epidermal growth factor represents an important class of the receptor tyrosine kinases playing the key role in the control of cell growth and differentiation in mammalian cells, as well as in the development of a number of pathological processes, including oncogene...
Gespeichert in:
Veröffentlicht in: | Biochemistry (Moscow). Supplement series A, Membrane and cell biology Membrane and cell biology, 2016-04, Vol.10 (2), p.142-149 |
---|---|
Hauptverfasser: | , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 149 |
---|---|
container_issue | 2 |
container_start_page | 142 |
container_title | Biochemistry (Moscow). Supplement series A, Membrane and cell biology |
container_volume | 10 |
creator | Bocharova, O. V. Bragin, P. E. Bocharov, E. V. Mineev, K. S. Goncharuk, S. A. Arseniev, A. S. |
description | The EGFR/HER receptor family of an epidermal growth factor represents an important class of the receptor tyrosine kinases playing the key role in the control of cell growth and differentiation in mammalian cells, as well as in the development of a number of pathological processes, including oncogenesis. Binding of a ligand to the extracellular domains initiates switching of the EGFR/HER receptor between the alternative dimeric states that causes the allosteric activation of kinase domains in cell cytoplasm. The transmembrane (TM) domain and adjacent flexible regions alternatively interacting with either membrane surface or kinase domains are directly involved in the complex conformational transition in EGFR/HERs. Here we report on a highly efficient system of the cell free production of the EGFR/HER TM domains with functionally important juxtamembrane (JM) regions for the investigation of the molecular basis of biochemical signal transduction across the cell membrane. To increase the efficiency of synthesis of the EGFR/HER TM-JM fragments of the receptors, we used two N-terminal expression tags, which significantly increased the protein yield. In the case of the TM-JM fragments of EGFR (residues 638–692) and HER2 (residues 644–700), the method allowed us to obtain milligram quantities of the
13
C,
15
N-labeled protein for structural and biophysical investigations in the membrane-mimicking environments using high-resolution heteronuclear NMR spectroscopy. |
doi_str_mv | 10.1134/S1990747816020045 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_1796450794</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>4088926971</sourcerecordid><originalsourceid>FETCH-LOGICAL-c268t-5722c92afbe1e8211f400712671fa7e6cbffe60b7c255dcd5095f5900ad489063</originalsourceid><addsrcrecordid>eNp1UdtKJDEQbURBV_0A3wL7artJTCedx2XwBsKCl-cmk66MGbvTbSWNzgfuf5lxdFhYfKriXOoUVUVxwugZY-fi1z3TmiqhaiYpp1RUO8XBGiqV0GJ326t6v_gR45JSeS6kPCj-zqDrSocABN5GhBj9EIgJLRkn9M5bk9bA4Eh6AuLQLHoIKX4BCBbGNCBJKxyiD0CefTARtoKLq8s74kzvu9UpsUNIxgcfFh9cQhNiD_08VyDt0GeOvPr09MEup7dktizCIu9xSlzOigknmyY0XW6n1kM8Kvac6SIcf9bD4vHy4mF2Xd7-ubqZ_b4tLZd1KivFudXcuDkwqDljTlCqGJeKOaNA2rlzIOlcWV5VrW0rqitXaUpNK2qdT3ZY_NzMHXF4mSCmZjlMGHJkw5SWoqJKi6xiG5XNN4kIrhnR9wZXDaPN-lvNf9_KHr7xxKwNC8B_Jn9regcXfptr</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1796450794</pqid></control><display><type>article</type><title>Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies</title><source>SpringerNature Journals</source><creator>Bocharova, O. V. ; Bragin, P. E. ; Bocharov, E. V. ; Mineev, K. S. ; Goncharuk, S. A. ; Arseniev, A. S.</creator><creatorcontrib>Bocharova, O. V. ; Bragin, P. E. ; Bocharov, E. V. ; Mineev, K. S. ; Goncharuk, S. A. ; Arseniev, A. S.</creatorcontrib><description>The EGFR/HER receptor family of an epidermal growth factor represents an important class of the receptor tyrosine kinases playing the key role in the control of cell growth and differentiation in mammalian cells, as well as in the development of a number of pathological processes, including oncogenesis. Binding of a ligand to the extracellular domains initiates switching of the EGFR/HER receptor between the alternative dimeric states that causes the allosteric activation of kinase domains in cell cytoplasm. The transmembrane (TM) domain and adjacent flexible regions alternatively interacting with either membrane surface or kinase domains are directly involved in the complex conformational transition in EGFR/HERs. Here we report on a highly efficient system of the cell free production of the EGFR/HER TM domains with functionally important juxtamembrane (JM) regions for the investigation of the molecular basis of biochemical signal transduction across the cell membrane. To increase the efficiency of synthesis of the EGFR/HER TM-JM fragments of the receptors, we used two N-terminal expression tags, which significantly increased the protein yield. In the case of the TM-JM fragments of EGFR (residues 638–692) and HER2 (residues 644–700), the method allowed us to obtain milligram quantities of the
13
C,
15
N-labeled protein for structural and biophysical investigations in the membrane-mimicking environments using high-resolution heteronuclear NMR spectroscopy.</description><identifier>ISSN: 1990-7478</identifier><identifier>EISSN: 1990-7494</identifier><identifier>DOI: 10.1134/S1990747816020045</identifier><language>eng</language><publisher>Moscow: Pleiades Publishing</publisher><subject>Biomedical and Life Sciences ; Cell Biology ; Cellular biology ; Kinases ; Life Sciences ; Membranes ; Spectrum analysis ; Studies</subject><ispartof>Biochemistry (Moscow). Supplement series A, Membrane and cell biology, 2016-04, Vol.10 (2), p.142-149</ispartof><rights>Pleiades Publishing, Ltd. 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c268t-5722c92afbe1e8211f400712671fa7e6cbffe60b7c255dcd5095f5900ad489063</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1134/S1990747816020045$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1134/S1990747816020045$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids></links><search><creatorcontrib>Bocharova, O. V.</creatorcontrib><creatorcontrib>Bragin, P. E.</creatorcontrib><creatorcontrib>Bocharov, E. V.</creatorcontrib><creatorcontrib>Mineev, K. S.</creatorcontrib><creatorcontrib>Goncharuk, S. A.</creatorcontrib><creatorcontrib>Arseniev, A. S.</creatorcontrib><title>Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies</title><title>Biochemistry (Moscow). Supplement series A, Membrane and cell biology</title><addtitle>Biochem. Moscow Suppl. Ser. A</addtitle><description>The EGFR/HER receptor family of an epidermal growth factor represents an important class of the receptor tyrosine kinases playing the key role in the control of cell growth and differentiation in mammalian cells, as well as in the development of a number of pathological processes, including oncogenesis. Binding of a ligand to the extracellular domains initiates switching of the EGFR/HER receptor between the alternative dimeric states that causes the allosteric activation of kinase domains in cell cytoplasm. The transmembrane (TM) domain and adjacent flexible regions alternatively interacting with either membrane surface or kinase domains are directly involved in the complex conformational transition in EGFR/HERs. Here we report on a highly efficient system of the cell free production of the EGFR/HER TM domains with functionally important juxtamembrane (JM) regions for the investigation of the molecular basis of biochemical signal transduction across the cell membrane. To increase the efficiency of synthesis of the EGFR/HER TM-JM fragments of the receptors, we used two N-terminal expression tags, which significantly increased the protein yield. In the case of the TM-JM fragments of EGFR (residues 638–692) and HER2 (residues 644–700), the method allowed us to obtain milligram quantities of the
13
C,
15
N-labeled protein for structural and biophysical investigations in the membrane-mimicking environments using high-resolution heteronuclear NMR spectroscopy.</description><subject>Biomedical and Life Sciences</subject><subject>Cell Biology</subject><subject>Cellular biology</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Membranes</subject><subject>Spectrum analysis</subject><subject>Studies</subject><issn>1990-7478</issn><issn>1990-7494</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1UdtKJDEQbURBV_0A3wL7artJTCedx2XwBsKCl-cmk66MGbvTbSWNzgfuf5lxdFhYfKriXOoUVUVxwugZY-fi1z3TmiqhaiYpp1RUO8XBGiqV0GJ326t6v_gR45JSeS6kPCj-zqDrSocABN5GhBj9EIgJLRkn9M5bk9bA4Eh6AuLQLHoIKX4BCBbGNCBJKxyiD0CefTARtoKLq8s74kzvu9UpsUNIxgcfFh9cQhNiD_08VyDt0GeOvPr09MEup7dktizCIu9xSlzOigknmyY0XW6n1kM8Kvac6SIcf9bD4vHy4mF2Xd7-ubqZ_b4tLZd1KivFudXcuDkwqDljTlCqGJeKOaNA2rlzIOlcWV5VrW0rqitXaUpNK2qdT3ZY_NzMHXF4mSCmZjlMGHJkw5SWoqJKi6xiG5XNN4kIrhnR9wZXDaPN-lvNf9_KHr7xxKwNC8B_Jn9regcXfptr</recordid><startdate>20160401</startdate><enddate>20160401</enddate><creator>Bocharova, O. V.</creator><creator>Bragin, P. E.</creator><creator>Bocharov, E. V.</creator><creator>Mineev, K. S.</creator><creator>Goncharuk, S. A.</creator><creator>Arseniev, A. S.</creator><general>Pleiades Publishing</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88I</scope><scope>8AO</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>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>LK8</scope><scope>M0S</scope><scope>M2P</scope><scope>M7P</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope></search><sort><creationdate>20160401</creationdate><title>Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies</title><author>Bocharova, O. V. ; Bragin, P. E. ; Bocharov, E. V. ; Mineev, K. S. ; Goncharuk, S. A. ; Arseniev, A. S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c268t-5722c92afbe1e8211f400712671fa7e6cbffe60b7c255dcd5095f5900ad489063</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Biomedical and Life Sciences</topic><topic>Cell Biology</topic><topic>Cellular biology</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Membranes</topic><topic>Spectrum analysis</topic><topic>Studies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bocharova, O. V.</creatorcontrib><creatorcontrib>Bragin, P. E.</creatorcontrib><creatorcontrib>Bocharov, E. V.</creatorcontrib><creatorcontrib>Mineev, K. S.</creatorcontrib><creatorcontrib>Goncharuk, S. A.</creatorcontrib><creatorcontrib>Arseniev, A. S.</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</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 Central UK/Ireland</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>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><jtitle>Biochemistry (Moscow). Supplement series A, Membrane and cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bocharova, O. V.</au><au>Bragin, P. E.</au><au>Bocharov, E. V.</au><au>Mineev, K. S.</au><au>Goncharuk, S. A.</au><au>Arseniev, A. S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies</atitle><jtitle>Biochemistry (Moscow). Supplement series A, Membrane and cell biology</jtitle><stitle>Biochem. Moscow Suppl. Ser. A</stitle><date>2016-04-01</date><risdate>2016</risdate><volume>10</volume><issue>2</issue><spage>142</spage><epage>149</epage><pages>142-149</pages><issn>1990-7478</issn><eissn>1990-7494</eissn><abstract>The EGFR/HER receptor family of an epidermal growth factor represents an important class of the receptor tyrosine kinases playing the key role in the control of cell growth and differentiation in mammalian cells, as well as in the development of a number of pathological processes, including oncogenesis. Binding of a ligand to the extracellular domains initiates switching of the EGFR/HER receptor between the alternative dimeric states that causes the allosteric activation of kinase domains in cell cytoplasm. The transmembrane (TM) domain and adjacent flexible regions alternatively interacting with either membrane surface or kinase domains are directly involved in the complex conformational transition in EGFR/HERs. Here we report on a highly efficient system of the cell free production of the EGFR/HER TM domains with functionally important juxtamembrane (JM) regions for the investigation of the molecular basis of biochemical signal transduction across the cell membrane. To increase the efficiency of synthesis of the EGFR/HER TM-JM fragments of the receptors, we used two N-terminal expression tags, which significantly increased the protein yield. In the case of the TM-JM fragments of EGFR (residues 638–692) and HER2 (residues 644–700), the method allowed us to obtain milligram quantities of the
13
C,
15
N-labeled protein for structural and biophysical investigations in the membrane-mimicking environments using high-resolution heteronuclear NMR spectroscopy.</abstract><cop>Moscow</cop><pub>Pleiades Publishing</pub><doi>10.1134/S1990747816020045</doi><tpages>8</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1990-7478 |
ispartof | Biochemistry (Moscow). Supplement series A, Membrane and cell biology, 2016-04, Vol.10 (2), p.142-149 |
issn | 1990-7478 1990-7494 |
language | eng |
recordid | cdi_proquest_journals_1796450794 |
source | SpringerNature Journals |
subjects | Biomedical and Life Sciences Cell Biology Cellular biology Kinases Life Sciences Membranes Spectrum analysis Studies |
title | Cell-free expression and purification of the fragments of the receptor tyrosine kinases of the EGFR family, containing the transmembrane domain with the juxtamembrane region, for structural studies |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T17%3A08%3A04IST&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=Cell-free%20expression%20and%20purification%20of%20the%20fragments%20of%20the%20receptor%20tyrosine%20kinases%20of%20the%20EGFR%20family,%20containing%20the%20transmembrane%20domain%20with%20the%20juxtamembrane%20region,%20for%20structural%20studies&rft.jtitle=Biochemistry%20(Moscow).%20Supplement%20series%20A,%20Membrane%20and%20cell%20biology&rft.au=Bocharova,%20O.%20V.&rft.date=2016-04-01&rft.volume=10&rft.issue=2&rft.spage=142&rft.epage=149&rft.pages=142-149&rft.issn=1990-7478&rft.eissn=1990-7494&rft_id=info:doi/10.1134/S1990747816020045&rft_dat=%3Cproquest_cross%3E4088926971%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=1796450794&rft_id=info:pmid/&rfr_iscdi=true |