The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25
The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatm...
Gespeichert in:
Veröffentlicht in: | International journal of biological macromolecules 1998-05, Vol.22 (3), p.163-173 |
---|---|
Hauptverfasser: | , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 173 |
---|---|
container_issue | 3 |
container_start_page | 163 |
container_title | International journal of biological macromolecules |
container_volume | 22 |
creator | Zavialov, Anton Benndorf, Rainer Ehrnsperger, Monika Zav’yalov, Vladimir Dudich, Igor Buchner, Johannes Gaestel, Matthias |
description | The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatment of the cells with diamide, arsenite, or hydrogen peroxide leads to an increase in Hsp25-dimerisation which can be blocked by simultaneous treatment with reducing agents. Recombinant Hsp25 was prepared in an oxidized dimeric (oxHsp25) and reduced monomeric (redHsp25) form. The two species were compared with regard to secondary structure, stability, oligomerization properties and their chaperone activity. It is demonstrated by CD measurements in the far UV region that there are no significant differences in the secondary structure and temperature- or pH-stability of oxHsp25 and redHsp25. However, according to CD measurements in the near UV region an increase in the asymmetry of the microenvironment of aromatic residues in oxHsp25 is observed. Furthermore, an increase in stability of the hydrophobic environment of the tryptophan residues mainly located in the N-terminal domain of the protein against urea denaturation is detected in oxHsp25. Both reduced and oxidized Hsp25 form oligomeric complexes of similar size and stability against detergents and both species prevent thermal aggregation of citrate synthase and assist significantly in oxaloacetic acid-induced refolding of the enzyme. Hence, the overall secondary structure, the degree of oligomerization and the chaperone activity of Hsp25 seem independent of the formation of the intermolecular disulfide bond and only the stability of the hydrophobic N-terminal part of the molecule is influenced by formation of this bound. The obtained data do not exclude the possible involvement of dimerization of this protein in other cellular functions, e.g. in intracellular sulfhydryl-buffering or in the protection of actin filaments from fragmentation upon oxidative stress. |
doi_str_mv | 10.1016/S0141-8130(98)00014-2 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_79975588</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0141813098000142</els_id><sourcerecordid>79975588</sourcerecordid><originalsourceid>FETCH-LOGICAL-c478t-42b9ebfc44664b92838403e08b15fb13c97dec4bdbc7e86fb4af7216e49493d83</originalsourceid><addsrcrecordid>eNqFkMtOHDEQRa0oiEyGfAKSV4gsOtjd7m57hRBKQiQkFgxrq22XNU567IkfSFnx63gesM2qdKtu1VUdhM4p-UYJHa4eCWW04bQjl4J_JaTKpv2AFpSPoqmy-4gW75ZP6HNKv2t36Ck_Radi6AkZ6QK9rNaAwVrQGQeLc1XOZ4ipqOJdxsalMltnAKvgDQ5-b0k5Fp1LnGY81a4tXmcXfJXbGLYQs4P0di5tpnnGa5gyTuug_-wsGZzHd2nb9mfoxE5zgi_HukRPP76vbu-a-4efv25v7hvNRp4b1ioBymrGhoEp0fKOM9IB4Yr2VtFOi9GAZsooPQIfrGKTHVs6ABNMdIZ3S3RxuFvT_xZIWW5c0jDPk4dQkhyFGPue74z9wahjSCmCldvoNlP8JymRO_ByD17uqErB5R68bOve-TGgqA2Y960j6Tq_PsyhfvnsIMqkHXgNxsUKX5rg_pPwCuhllH0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>79975588</pqid></control><display><type>article</type><title>The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Zavialov, Anton ; Benndorf, Rainer ; Ehrnsperger, Monika ; Zav’yalov, Vladimir ; Dudich, Igor ; Buchner, Johannes ; Gaestel, Matthias</creator><creatorcontrib>Zavialov, Anton ; Benndorf, Rainer ; Ehrnsperger, Monika ; Zav’yalov, Vladimir ; Dudich, Igor ; Buchner, Johannes ; Gaestel, Matthias</creatorcontrib><description>The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatment of the cells with diamide, arsenite, or hydrogen peroxide leads to an increase in Hsp25-dimerisation which can be blocked by simultaneous treatment with reducing agents. Recombinant Hsp25 was prepared in an oxidized dimeric (oxHsp25) and reduced monomeric (redHsp25) form. The two species were compared with regard to secondary structure, stability, oligomerization properties and their chaperone activity. It is demonstrated by CD measurements in the far UV region that there are no significant differences in the secondary structure and temperature- or pH-stability of oxHsp25 and redHsp25. However, according to CD measurements in the near UV region an increase in the asymmetry of the microenvironment of aromatic residues in oxHsp25 is observed. Furthermore, an increase in stability of the hydrophobic environment of the tryptophan residues mainly located in the N-terminal domain of the protein against urea denaturation is detected in oxHsp25. Both reduced and oxidized Hsp25 form oligomeric complexes of similar size and stability against detergents and both species prevent thermal aggregation of citrate synthase and assist significantly in oxaloacetic acid-induced refolding of the enzyme. Hence, the overall secondary structure, the degree of oligomerization and the chaperone activity of Hsp25 seem independent of the formation of the intermolecular disulfide bond and only the stability of the hydrophobic N-terminal part of the molecule is influenced by formation of this bound. The obtained data do not exclude the possible involvement of dimerization of this protein in other cellular functions, e.g. in intracellular sulfhydryl-buffering or in the protection of actin filaments from fragmentation upon oxidative stress.</description><identifier>ISSN: 0141-8130</identifier><identifier>EISSN: 1879-0003</identifier><identifier>DOI: 10.1016/S0141-8130(98)00014-2</identifier><identifier>PMID: 9650071</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Amino Acid Sequence ; Animals ; Chaperone ; Circular Dichroism ; Dimerization ; Dimerzation ; Disulfides - chemistry ; Drug Stability ; Heat-Shock Proteins - chemistry ; Heat-Shock Proteins - genetics ; Heat-Shock Proteins - physiology ; HSP27 Heat-Shock Proteins ; Humans ; In Vitro Techniques ; Mice ; Molecular Chaperones - chemistry ; Molecular Chaperones - genetics ; Molecular Chaperones - physiology ; Molecular Sequence Data ; Neoplasm Proteins - chemistry ; Neoplasm Proteins - genetics ; Neoplasm Proteins - physiology ; Oligomerization ; Oxidation-Reduction ; Oxidative Stress ; Protein Conformation ; Protein Structure, Secondary ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Sequence Homology, Amino Acid ; Tumor Cells, Cultured</subject><ispartof>International journal of biological macromolecules, 1998-05, Vol.22 (3), p.163-173</ispartof><rights>1998 Elsevier Science B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-42b9ebfc44664b92838403e08b15fb13c97dec4bdbc7e86fb4af7216e49493d83</citedby><cites>FETCH-LOGICAL-c478t-42b9ebfc44664b92838403e08b15fb13c97dec4bdbc7e86fb4af7216e49493d83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0141-8130(98)00014-2$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9650071$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zavialov, Anton</creatorcontrib><creatorcontrib>Benndorf, Rainer</creatorcontrib><creatorcontrib>Ehrnsperger, Monika</creatorcontrib><creatorcontrib>Zav’yalov, Vladimir</creatorcontrib><creatorcontrib>Dudich, Igor</creatorcontrib><creatorcontrib>Buchner, Johannes</creatorcontrib><creatorcontrib>Gaestel, Matthias</creatorcontrib><title>The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25</title><title>International journal of biological macromolecules</title><addtitle>Int J Biol Macromol</addtitle><description>The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatment of the cells with diamide, arsenite, or hydrogen peroxide leads to an increase in Hsp25-dimerisation which can be blocked by simultaneous treatment with reducing agents. Recombinant Hsp25 was prepared in an oxidized dimeric (oxHsp25) and reduced monomeric (redHsp25) form. The two species were compared with regard to secondary structure, stability, oligomerization properties and their chaperone activity. It is demonstrated by CD measurements in the far UV region that there are no significant differences in the secondary structure and temperature- or pH-stability of oxHsp25 and redHsp25. However, according to CD measurements in the near UV region an increase in the asymmetry of the microenvironment of aromatic residues in oxHsp25 is observed. Furthermore, an increase in stability of the hydrophobic environment of the tryptophan residues mainly located in the N-terminal domain of the protein against urea denaturation is detected in oxHsp25. Both reduced and oxidized Hsp25 form oligomeric complexes of similar size and stability against detergents and both species prevent thermal aggregation of citrate synthase and assist significantly in oxaloacetic acid-induced refolding of the enzyme. Hence, the overall secondary structure, the degree of oligomerization and the chaperone activity of Hsp25 seem independent of the formation of the intermolecular disulfide bond and only the stability of the hydrophobic N-terminal part of the molecule is influenced by formation of this bound. The obtained data do not exclude the possible involvement of dimerization of this protein in other cellular functions, e.g. in intracellular sulfhydryl-buffering or in the protection of actin filaments from fragmentation upon oxidative stress.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Chaperone</subject><subject>Circular Dichroism</subject><subject>Dimerization</subject><subject>Dimerzation</subject><subject>Disulfides - chemistry</subject><subject>Drug Stability</subject><subject>Heat-Shock Proteins - chemistry</subject><subject>Heat-Shock Proteins - genetics</subject><subject>Heat-Shock Proteins - physiology</subject><subject>HSP27 Heat-Shock Proteins</subject><subject>Humans</subject><subject>In Vitro Techniques</subject><subject>Mice</subject><subject>Molecular Chaperones - chemistry</subject><subject>Molecular Chaperones - genetics</subject><subject>Molecular Chaperones - physiology</subject><subject>Molecular Sequence Data</subject><subject>Neoplasm Proteins - chemistry</subject><subject>Neoplasm Proteins - genetics</subject><subject>Neoplasm Proteins - physiology</subject><subject>Oligomerization</subject><subject>Oxidation-Reduction</subject><subject>Oxidative Stress</subject><subject>Protein Conformation</subject><subject>Protein Structure, Secondary</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Sequence Homology, Amino Acid</subject><subject>Tumor Cells, Cultured</subject><issn>0141-8130</issn><issn>1879-0003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOHDEQRa0oiEyGfAKSV4gsOtjd7m57hRBKQiQkFgxrq22XNU567IkfSFnx63gesM2qdKtu1VUdhM4p-UYJHa4eCWW04bQjl4J_JaTKpv2AFpSPoqmy-4gW75ZP6HNKv2t36Ck_Radi6AkZ6QK9rNaAwVrQGQeLc1XOZ4ipqOJdxsalMltnAKvgDQ5-b0k5Fp1LnGY81a4tXmcXfJXbGLYQs4P0di5tpnnGa5gyTuug_-wsGZzHd2nb9mfoxE5zgi_HukRPP76vbu-a-4efv25v7hvNRp4b1ioBymrGhoEp0fKOM9IB4Yr2VtFOi9GAZsooPQIfrGKTHVs6ABNMdIZ3S3RxuFvT_xZIWW5c0jDPk4dQkhyFGPue74z9wahjSCmCldvoNlP8JymRO_ByD17uqErB5R68bOve-TGgqA2Y960j6Tq_PsyhfvnsIMqkHXgNxsUKX5rg_pPwCuhllH0</recordid><startdate>19980501</startdate><enddate>19980501</enddate><creator>Zavialov, Anton</creator><creator>Benndorf, Rainer</creator><creator>Ehrnsperger, Monika</creator><creator>Zav’yalov, Vladimir</creator><creator>Dudich, Igor</creator><creator>Buchner, Johannes</creator><creator>Gaestel, Matthias</creator><general>Elsevier B.V</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>19980501</creationdate><title>The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25</title><author>Zavialov, Anton ; Benndorf, Rainer ; Ehrnsperger, Monika ; Zav’yalov, Vladimir ; Dudich, Igor ; Buchner, Johannes ; Gaestel, Matthias</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-42b9ebfc44664b92838403e08b15fb13c97dec4bdbc7e86fb4af7216e49493d83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Chaperone</topic><topic>Circular Dichroism</topic><topic>Dimerization</topic><topic>Dimerzation</topic><topic>Disulfides - chemistry</topic><topic>Drug Stability</topic><topic>Heat-Shock Proteins - chemistry</topic><topic>Heat-Shock Proteins - genetics</topic><topic>Heat-Shock Proteins - physiology</topic><topic>HSP27 Heat-Shock Proteins</topic><topic>Humans</topic><topic>In Vitro Techniques</topic><topic>Mice</topic><topic>Molecular Chaperones - chemistry</topic><topic>Molecular Chaperones - genetics</topic><topic>Molecular Chaperones - physiology</topic><topic>Molecular Sequence Data</topic><topic>Neoplasm Proteins - chemistry</topic><topic>Neoplasm Proteins - genetics</topic><topic>Neoplasm Proteins - physiology</topic><topic>Oligomerization</topic><topic>Oxidation-Reduction</topic><topic>Oxidative Stress</topic><topic>Protein Conformation</topic><topic>Protein Structure, Secondary</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Sequence Homology, Amino Acid</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zavialov, Anton</creatorcontrib><creatorcontrib>Benndorf, Rainer</creatorcontrib><creatorcontrib>Ehrnsperger, Monika</creatorcontrib><creatorcontrib>Zav’yalov, Vladimir</creatorcontrib><creatorcontrib>Dudich, Igor</creatorcontrib><creatorcontrib>Buchner, Johannes</creatorcontrib><creatorcontrib>Gaestel, Matthias</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>International journal of biological macromolecules</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zavialov, Anton</au><au>Benndorf, Rainer</au><au>Ehrnsperger, Monika</au><au>Zav’yalov, Vladimir</au><au>Dudich, Igor</au><au>Buchner, Johannes</au><au>Gaestel, Matthias</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25</atitle><jtitle>International journal of biological macromolecules</jtitle><addtitle>Int J Biol Macromol</addtitle><date>1998-05-01</date><risdate>1998</risdate><volume>22</volume><issue>3</issue><spage>163</spage><epage>173</epage><pages>163-173</pages><issn>0141-8130</issn><eissn>1879-0003</eissn><abstract>The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatment of the cells with diamide, arsenite, or hydrogen peroxide leads to an increase in Hsp25-dimerisation which can be blocked by simultaneous treatment with reducing agents. Recombinant Hsp25 was prepared in an oxidized dimeric (oxHsp25) and reduced monomeric (redHsp25) form. The two species were compared with regard to secondary structure, stability, oligomerization properties and their chaperone activity. It is demonstrated by CD measurements in the far UV region that there are no significant differences in the secondary structure and temperature- or pH-stability of oxHsp25 and redHsp25. However, according to CD measurements in the near UV region an increase in the asymmetry of the microenvironment of aromatic residues in oxHsp25 is observed. Furthermore, an increase in stability of the hydrophobic environment of the tryptophan residues mainly located in the N-terminal domain of the protein against urea denaturation is detected in oxHsp25. Both reduced and oxidized Hsp25 form oligomeric complexes of similar size and stability against detergents and both species prevent thermal aggregation of citrate synthase and assist significantly in oxaloacetic acid-induced refolding of the enzyme. Hence, the overall secondary structure, the degree of oligomerization and the chaperone activity of Hsp25 seem independent of the formation of the intermolecular disulfide bond and only the stability of the hydrophobic N-terminal part of the molecule is influenced by formation of this bound. The obtained data do not exclude the possible involvement of dimerization of this protein in other cellular functions, e.g. in intracellular sulfhydryl-buffering or in the protection of actin filaments from fragmentation upon oxidative stress.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>9650071</pmid><doi>10.1016/S0141-8130(98)00014-2</doi><tpages>11</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0141-8130 |
ispartof | International journal of biological macromolecules, 1998-05, Vol.22 (3), p.163-173 |
issn | 0141-8130 1879-0003 |
language | eng |
recordid | cdi_proquest_miscellaneous_79975588 |
source | MEDLINE; Access via ScienceDirect (Elsevier) |
subjects | Amino Acid Sequence Animals Chaperone Circular Dichroism Dimerization Dimerzation Disulfides - chemistry Drug Stability Heat-Shock Proteins - chemistry Heat-Shock Proteins - genetics Heat-Shock Proteins - physiology HSP27 Heat-Shock Proteins Humans In Vitro Techniques Mice Molecular Chaperones - chemistry Molecular Chaperones - genetics Molecular Chaperones - physiology Molecular Sequence Data Neoplasm Proteins - chemistry Neoplasm Proteins - genetics Neoplasm Proteins - physiology Oligomerization Oxidation-Reduction Oxidative Stress Protein Conformation Protein Structure, Secondary Recombinant Proteins - chemistry Recombinant Proteins - genetics Sequence Homology, Amino Acid Tumor Cells, Cultured |
title | The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25 |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-15T22%3A16%3A09IST&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=The%20effect%20of%20the%20intersubunit%20disulfide%20bond%20on%20the%20structural%20and%20functional%20properties%20of%20the%20small%20heat%20shock%20protein%20Hsp25&rft.jtitle=International%20journal%20of%20biological%20macromolecules&rft.au=Zavialov,%20Anton&rft.date=1998-05-01&rft.volume=22&rft.issue=3&rft.spage=163&rft.epage=173&rft.pages=163-173&rft.issn=0141-8130&rft.eissn=1879-0003&rft_id=info:doi/10.1016/S0141-8130(98)00014-2&rft_dat=%3Cproquest_cross%3E79975588%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=79975588&rft_id=info:pmid/9650071&rft_els_id=S0141813098000142&rfr_iscdi=true |