The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria
Bax is a major player in the apoptotic process, being at the core of the mitochondria permeabilization events. In spite of the major recent advances in the knowledge of Bax organization within the membrane, the precise behavior of the C-terminal helix α9 remains elusive, since it was absent from the...
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| Veröffentlicht in: | Biochimica et biophysica acta. Biomembranes 2017-06, Vol.1859 (6), p.1144-1155 |
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| creator | Simonyan, Lilit Légiot, Alexandre Lascu, Ioan Durand, Grégory Giraud, Marie-France Gonzalez, Cécile Manon, Stéphen |
| description | Bax is a major player in the apoptotic process, being at the core of the mitochondria permeabilization events. In spite of the major recent advances in the knowledge of Bax organization within the membrane, the precise behavior of the C-terminal helix α9 remains elusive, since it was absent from the resolved structure of active Bax. The Proline 168 (P168) residue, located in the short loop between α8 and α9, has been the target of site-directed mutagenesis experiments, with conflicting results. We have produced and purified a recombinant mutant Bax-P168A, and we have compared its behavior with that of wild-type Bax in a series of tests on Large Unilamellar Vesicles (LUVs) and isolated mitochondria. We conclude that Bax-P168A had a greater ability to oligomerize and bind to membranes. Bax-P168A was not more efficient than wild-type Bax to permeabilize liposomes to small molecules but was more prone to release cytochrome c from mitochondria.
[Display omitted]
•Biochemical effect of the mutation P168A of Bax was studied.•Bax-P168A is prone to oligomerize more spontaneously than BaxWT.•It is not significantly more active than BaxWT to permeabilize liposomes.•It is more active than BaxWT to permeabilize mitochondria. |
| doi_str_mv | 10.1016/j.bbamem.2017.03.010 |
| format | Article |
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[Display omitted]
•Biochemical effect of the mutation P168A of Bax was studied.•Bax-P168A is prone to oligomerize more spontaneously than BaxWT.•It is not significantly more active than BaxWT to permeabilize liposomes.•It is more active than BaxWT to permeabilize mitochondria.</description><identifier>ISSN: 0005-2736</identifier><identifier>EISSN: 1879-2642</identifier><identifier>DOI: 10.1016/j.bbamem.2017.03.010</identifier><identifier>PMID: 28322731</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Alanine ; Alanine - chemistry ; Alanine - metabolism ; Amino Acid Substitution ; Bax ; bcl-2-Associated X Protein ; bcl-2-Associated X Protein - chemistry ; bcl-2-Associated X Protein - genetics ; bcl-2-Associated X Protein - metabolism ; Cell-free assays ; Cell-free expression ; Chemical Sciences ; Cloning, Molecular ; Cytochromes c ; Cytochromes c - secretion ; Gene Expression ; HCT116 Cells ; Humans ; Liposomes ; Liposomes - chemistry ; Liposomes - metabolism ; LUV ; Mitochondria ; Mitochondria - chemistry ; Mitochondria - metabolism ; Mutation ; Permeability ; Proline ; Proline - chemistry ; Proline - metabolism ; Protein Binding ; Protein Multimerization ; Protein Structure, Secondary ; Recombinant Proteins ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - metabolism ; Unilamellar Liposomes ; Unilamellar Liposomes - chemistry ; Unilamellar Liposomes - metabolism</subject><ispartof>Biochimica et biophysica acta. Biomembranes, 2017-06, Vol.1859 (6), p.1144-1155</ispartof><rights>2017 Elsevier B.V.</rights><rights>Copyright © 2017 Elsevier B.V. All rights reserved.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c442t-240430297af82207e478a34b5fadaf1084ce1c96b224030eb23a4214b12f7b1f3</citedby><cites>FETCH-LOGICAL-c442t-240430297af82207e478a34b5fadaf1084ce1c96b224030eb23a4214b12f7b1f3</cites><orcidid>0000-0001-8623-3443 ; 0000-0002-6238-667X ; 0000-0001-6680-2821</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbamem.2017.03.010$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28322731$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.umontpellier.fr/hal-03559837$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Simonyan, Lilit</creatorcontrib><creatorcontrib>Légiot, Alexandre</creatorcontrib><creatorcontrib>Lascu, Ioan</creatorcontrib><creatorcontrib>Durand, Grégory</creatorcontrib><creatorcontrib>Giraud, Marie-France</creatorcontrib><creatorcontrib>Gonzalez, Cécile</creatorcontrib><creatorcontrib>Manon, Stéphen</creatorcontrib><title>The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria</title><title>Biochimica et biophysica acta. Biomembranes</title><addtitle>Biochim Biophys Acta Biomembr</addtitle><description>Bax is a major player in the apoptotic process, being at the core of the mitochondria permeabilization events. In spite of the major recent advances in the knowledge of Bax organization within the membrane, the precise behavior of the C-terminal helix α9 remains elusive, since it was absent from the resolved structure of active Bax. The Proline 168 (P168) residue, located in the short loop between α8 and α9, has been the target of site-directed mutagenesis experiments, with conflicting results. We have produced and purified a recombinant mutant Bax-P168A, and we have compared its behavior with that of wild-type Bax in a series of tests on Large Unilamellar Vesicles (LUVs) and isolated mitochondria. We conclude that Bax-P168A had a greater ability to oligomerize and bind to membranes. Bax-P168A was not more efficient than wild-type Bax to permeabilize liposomes to small molecules but was more prone to release cytochrome c from mitochondria.
[Display omitted]
•Biochemical effect of the mutation P168A of Bax was studied.•Bax-P168A is prone to oligomerize more spontaneously than BaxWT.•It is not significantly more active than BaxWT to permeabilize liposomes.•It is more active than BaxWT to permeabilize mitochondria.</description><subject>Alanine</subject><subject>Alanine - chemistry</subject><subject>Alanine - metabolism</subject><subject>Amino Acid Substitution</subject><subject>Bax</subject><subject>bcl-2-Associated X Protein</subject><subject>bcl-2-Associated X Protein - chemistry</subject><subject>bcl-2-Associated X Protein - genetics</subject><subject>bcl-2-Associated X Protein - metabolism</subject><subject>Cell-free assays</subject><subject>Cell-free expression</subject><subject>Chemical Sciences</subject><subject>Cloning, Molecular</subject><subject>Cytochromes c</subject><subject>Cytochromes c - secretion</subject><subject>Gene Expression</subject><subject>HCT116 Cells</subject><subject>Humans</subject><subject>Liposomes</subject><subject>Liposomes - chemistry</subject><subject>Liposomes - metabolism</subject><subject>LUV</subject><subject>Mitochondria</subject><subject>Mitochondria - chemistry</subject><subject>Mitochondria - metabolism</subject><subject>Mutation</subject><subject>Permeability</subject><subject>Proline</subject><subject>Proline - chemistry</subject><subject>Proline - metabolism</subject><subject>Protein Binding</subject><subject>Protein Multimerization</subject><subject>Protein Structure, Secondary</subject><subject>Recombinant Proteins</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Unilamellar Liposomes</subject><subject>Unilamellar Liposomes - chemistry</subject><subject>Unilamellar Liposomes - metabolism</subject><issn>0005-2736</issn><issn>1879-2642</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kEtvEzEURi0EakPpP0DIWxYzXD_mkQ1SqVqKFAkWLVvL9lwTR5lxZTvp49fjZKDLLqwrffccW_4I-cigZsDaL5vaGD3iWHNgXQ2iBgZvyIL13bLireRvyQIAmop3oj0l71PaQNEkb07IKe8FLzlbkHy7Rpp2JmWfd9mHiQZHf8Ww9RNS1vbU6X2IiX7Tj7SEf8KI0T_rI6mngRZv3G11xkR9LmfKGLU9rh98XtPV3e90BEefg12HaYhefyDvnN4mPP83z8jd9dXt5U21-vn9x-XFqrJS8lxxCVIAX3ba9ZxDh7LrtZCmcXrQjkEvLTK7bA0vpAA0XGjJmTSMu84wJ87I5_netd6q--hHHZ9U0F7dXKzUIQPRNMtedHtWWDmzNoaUIroXgYE6FK42ai5cHQovqiqFF-3TrN3vzIjDi_S_4QJ8nQEsH917jCpZj5PFwUe0WQ3Bv_7CX59vk3M</recordid><startdate>201706</startdate><enddate>201706</enddate><creator>Simonyan, Lilit</creator><creator>Légiot, Alexandre</creator><creator>Lascu, Ioan</creator><creator>Durand, Grégory</creator><creator>Giraud, Marie-France</creator><creator>Gonzalez, Cécile</creator><creator>Manon, Stéphen</creator><general>Elsevier B.V</general><general>Elsevier</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>1XC</scope><orcidid>https://orcid.org/0000-0001-8623-3443</orcidid><orcidid>https://orcid.org/0000-0002-6238-667X</orcidid><orcidid>https://orcid.org/0000-0001-6680-2821</orcidid></search><sort><creationdate>201706</creationdate><title>The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria</title><author>Simonyan, Lilit ; Légiot, Alexandre ; Lascu, Ioan ; Durand, Grégory ; Giraud, Marie-France ; Gonzalez, Cécile ; Manon, Stéphen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c442t-240430297af82207e478a34b5fadaf1084ce1c96b224030eb23a4214b12f7b1f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Alanine</topic><topic>Alanine - chemistry</topic><topic>Alanine - metabolism</topic><topic>Amino Acid Substitution</topic><topic>Bax</topic><topic>bcl-2-Associated X Protein</topic><topic>bcl-2-Associated X Protein - chemistry</topic><topic>bcl-2-Associated X Protein - genetics</topic><topic>bcl-2-Associated X Protein - metabolism</topic><topic>Cell-free assays</topic><topic>Cell-free expression</topic><topic>Chemical Sciences</topic><topic>Cloning, Molecular</topic><topic>Cytochromes c</topic><topic>Cytochromes c - secretion</topic><topic>Gene Expression</topic><topic>HCT116 Cells</topic><topic>Humans</topic><topic>Liposomes</topic><topic>Liposomes - chemistry</topic><topic>Liposomes - metabolism</topic><topic>LUV</topic><topic>Mitochondria</topic><topic>Mitochondria - chemistry</topic><topic>Mitochondria - metabolism</topic><topic>Mutation</topic><topic>Permeability</topic><topic>Proline</topic><topic>Proline - chemistry</topic><topic>Proline - metabolism</topic><topic>Protein Binding</topic><topic>Protein Multimerization</topic><topic>Protein Structure, Secondary</topic><topic>Recombinant Proteins</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Unilamellar Liposomes</topic><topic>Unilamellar Liposomes - chemistry</topic><topic>Unilamellar Liposomes - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Simonyan, Lilit</creatorcontrib><creatorcontrib>Légiot, Alexandre</creatorcontrib><creatorcontrib>Lascu, Ioan</creatorcontrib><creatorcontrib>Durand, Grégory</creatorcontrib><creatorcontrib>Giraud, Marie-France</creatorcontrib><creatorcontrib>Gonzalez, Cécile</creatorcontrib><creatorcontrib>Manon, Stéphen</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>Hyper Article en Ligne (HAL)</collection><jtitle>Biochimica et biophysica acta. Biomembranes</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Simonyan, Lilit</au><au>Légiot, Alexandre</au><au>Lascu, Ioan</au><au>Durand, Grégory</au><au>Giraud, Marie-France</au><au>Gonzalez, Cécile</au><au>Manon, Stéphen</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria</atitle><jtitle>Biochimica et biophysica acta. Biomembranes</jtitle><addtitle>Biochim Biophys Acta Biomembr</addtitle><date>2017-06</date><risdate>2017</risdate><volume>1859</volume><issue>6</issue><spage>1144</spage><epage>1155</epage><pages>1144-1155</pages><issn>0005-2736</issn><eissn>1879-2642</eissn><abstract>Bax is a major player in the apoptotic process, being at the core of the mitochondria permeabilization events. In spite of the major recent advances in the knowledge of Bax organization within the membrane, the precise behavior of the C-terminal helix α9 remains elusive, since it was absent from the resolved structure of active Bax. The Proline 168 (P168) residue, located in the short loop between α8 and α9, has been the target of site-directed mutagenesis experiments, with conflicting results. We have produced and purified a recombinant mutant Bax-P168A, and we have compared its behavior with that of wild-type Bax in a series of tests on Large Unilamellar Vesicles (LUVs) and isolated mitochondria. We conclude that Bax-P168A had a greater ability to oligomerize and bind to membranes. Bax-P168A was not more efficient than wild-type Bax to permeabilize liposomes to small molecules but was more prone to release cytochrome c from mitochondria.
[Display omitted]
•Biochemical effect of the mutation P168A of Bax was studied.•Bax-P168A is prone to oligomerize more spontaneously than BaxWT.•It is not significantly more active than BaxWT to permeabilize liposomes.•It is more active than BaxWT to permeabilize mitochondria.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>28322731</pmid><doi>10.1016/j.bbamem.2017.03.010</doi><tpages>12</tpages><orcidid>https://orcid.org/0000-0001-8623-3443</orcidid><orcidid>https://orcid.org/0000-0002-6238-667X</orcidid><orcidid>https://orcid.org/0000-0001-6680-2821</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Biochimica et biophysica acta. Biomembranes, 2017-06, Vol.1859 (6), p.1144-1155 |
| issn | 0005-2736 1879-2642 |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present); EZB-FREE-00999 freely available EZB journals |
| subjects | Alanine Alanine - chemistry Alanine - metabolism Amino Acid Substitution Bax bcl-2-Associated X Protein bcl-2-Associated X Protein - chemistry bcl-2-Associated X Protein - genetics bcl-2-Associated X Protein - metabolism Cell-free assays Cell-free expression Chemical Sciences Cloning, Molecular Cytochromes c Cytochromes c - secretion Gene Expression HCT116 Cells Humans Liposomes Liposomes - chemistry Liposomes - metabolism LUV Mitochondria Mitochondria - chemistry Mitochondria - metabolism Mutation Permeability Proline Proline - chemistry Proline - metabolism Protein Binding Protein Multimerization Protein Structure, Secondary Recombinant Proteins Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Unilamellar Liposomes Unilamellar Liposomes - chemistry Unilamellar Liposomes - metabolism |
| title | The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria |
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