Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c
A cell-free system based on cytosols of normally growing cells is established that reproduces aspects of the apoptotic program in vitro. The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption s...
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| Veröffentlicht in: | Cell 1996-07, Vol.86 (1), p.147-157 |
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| creator | Liu, Xuesong Kim, Caryn Naekyung Yang, Jie Jemmerson, Ronald Wang, Xiaodong |
| description | A cell-free system based on cytosols of normally growing cells is established that reproduces aspects of the apoptotic program in vitro. The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption spectrum and protein sequence revealed that this protein is cytochrome c. Elimination of cytochrome c from cytosol by immunodepletion, or inclusion of sucrose to stabilize mitochondria during cytosol preparation, diminished the apoptotic activity. Adding back cytochrome c to the cytochrome c–depleted extracts restored their apoptotic activity. Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c. |
| doi_str_mv | 10.1016/S0092-8674(00)80085-9 |
| format | Article |
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The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption spectrum and protein sequence revealed that this protein is cytochrome c. Elimination of cytochrome c from cytosol by immunodepletion, or inclusion of sucrose to stabilize mitochondria during cytosol preparation, diminished the apoptotic activity. Adding back cytochrome c to the cytochrome c–depleted extracts restored their apoptotic activity. Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c.</description><identifier>ISSN: 0092-8674</identifier><identifier>EISSN: 1097-4172</identifier><identifier>DOI: 10.1016/S0092-8674(00)80085-9</identifier><identifier>PMID: 8689682</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Adenosine Triphosphate - metabolism ; Amino Acid Sequence ; Antibodies, Monoclonal ; Apoptosis - drug effects ; Caspase 3 ; Caspases ; Cell Extracts - chemistry ; Cell Extracts - pharmacology ; Cell-Free System ; Cysteine Endopeptidases - metabolism ; Cytochrome c Group - immunology ; Cytochrome c Group - isolation & purification ; Cytochrome c Group - metabolism ; Cytosol - chemistry ; Cytosol - metabolism ; DNA - drug effects ; DNA - metabolism ; Enzyme Precursors - metabolism ; HeLa Cells - cytology ; HeLa Cells - drug effects ; HeLa Cells - enzymology ; Humans ; Molecular Sequence Data</subject><ispartof>Cell, 1996-07, Vol.86 (1), p.147-157</ispartof><rights>1996 Cell Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c556t-d73494c9e34f476ffd7241e61296e168eaa6ab497f241c10cab75e46f81dbc453</citedby><cites>FETCH-LOGICAL-c556t-d73494c9e34f476ffd7241e61296e168eaa6ab497f241c10cab75e46f81dbc453</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0092867400800859$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8689682$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Xuesong</creatorcontrib><creatorcontrib>Kim, Caryn Naekyung</creatorcontrib><creatorcontrib>Yang, Jie</creatorcontrib><creatorcontrib>Jemmerson, Ronald</creatorcontrib><creatorcontrib>Wang, Xiaodong</creatorcontrib><title>Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c</title><title>Cell</title><addtitle>Cell</addtitle><description>A cell-free system based on cytosols of normally growing cells is established that reproduces aspects of the apoptotic program in vitro. The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption spectrum and protein sequence revealed that this protein is cytochrome c. Elimination of cytochrome c from cytosol by immunodepletion, or inclusion of sucrose to stabilize mitochondria during cytosol preparation, diminished the apoptotic activity. Adding back cytochrome c to the cytochrome c–depleted extracts restored their apoptotic activity. Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Antibodies, Monoclonal</subject><subject>Apoptosis - drug effects</subject><subject>Caspase 3</subject><subject>Caspases</subject><subject>Cell Extracts - chemistry</subject><subject>Cell Extracts - pharmacology</subject><subject>Cell-Free System</subject><subject>Cysteine Endopeptidases - metabolism</subject><subject>Cytochrome c Group - immunology</subject><subject>Cytochrome c Group - isolation & purification</subject><subject>Cytochrome c Group - metabolism</subject><subject>Cytosol - chemistry</subject><subject>Cytosol - metabolism</subject><subject>DNA - drug effects</subject><subject>DNA - metabolism</subject><subject>Enzyme Precursors - metabolism</subject><subject>HeLa Cells - cytology</subject><subject>HeLa Cells - drug effects</subject><subject>HeLa Cells - enzymology</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><issn>0092-8674</issn><issn>1097-4172</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1rGzEQhkVpSB2nPyGgU2kOm0hrffZSjMkXGBLycepByNKoVfGuHElbmn-fdWxyzWlg5nlnhgehE0rOKKHi_IEQ3TZKSPadkFNFiOKN_oQmlGjZMCrbz2jyjnxBR6X8JVuI80N0qITSQrUT9Oum94OrMfU4BTzfpE1NNTp8l9PvbDsce7yA9bq5zAD44n_N1tXyA9_D8xAzdNBXHFLGfv54h23v8eKlJvcnpw6wO0YHwa4LfN3XKXq6vHhcXDfL26ubxXzZOM5FbbycMc2chhkLTIoQvGwZBUFbLYAKBdYKu2JahrHtKHF2JTkwERT1K8f4bIq-7fZucnoeoFTTxeLGr20PaShGKqoonYkPQcqFYJTIEeQ70OVUSoZgNjl2Nr8YSszWvnmzb7ZqDSHmzb7RY-5kf2BYdeDfU3vd4_znbg6jjn8RsikuQu_AjzJdNT7FDy68Apnrk5o</recordid><startdate>19960712</startdate><enddate>19960712</enddate><creator>Liu, Xuesong</creator><creator>Kim, Caryn Naekyung</creator><creator>Yang, Jie</creator><creator>Jemmerson, Ronald</creator><creator>Wang, Xiaodong</creator><general>Elsevier Inc</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>7TM</scope><scope>7X8</scope></search><sort><creationdate>19960712</creationdate><title>Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c</title><author>Liu, Xuesong ; Kim, Caryn Naekyung ; Yang, Jie ; Jemmerson, Ronald ; Wang, Xiaodong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c556t-d73494c9e34f476ffd7241e61296e168eaa6ab497f241c10cab75e46f81dbc453</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Antibodies, Monoclonal</topic><topic>Apoptosis - drug effects</topic><topic>Caspase 3</topic><topic>Caspases</topic><topic>Cell Extracts - chemistry</topic><topic>Cell Extracts - pharmacology</topic><topic>Cell-Free System</topic><topic>Cysteine Endopeptidases - metabolism</topic><topic>Cytochrome c Group - immunology</topic><topic>Cytochrome c Group - isolation & purification</topic><topic>Cytochrome c Group - metabolism</topic><topic>Cytosol - chemistry</topic><topic>Cytosol - metabolism</topic><topic>DNA - drug effects</topic><topic>DNA - metabolism</topic><topic>Enzyme Precursors - metabolism</topic><topic>HeLa Cells - cytology</topic><topic>HeLa Cells - drug effects</topic><topic>HeLa Cells - enzymology</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Xuesong</creatorcontrib><creatorcontrib>Kim, Caryn Naekyung</creatorcontrib><creatorcontrib>Yang, Jie</creatorcontrib><creatorcontrib>Jemmerson, Ronald</creatorcontrib><creatorcontrib>Wang, Xiaodong</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>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cell</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Xuesong</au><au>Kim, Caryn Naekyung</au><au>Yang, Jie</au><au>Jemmerson, Ronald</au><au>Wang, Xiaodong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c</atitle><jtitle>Cell</jtitle><addtitle>Cell</addtitle><date>1996-07-12</date><risdate>1996</risdate><volume>86</volume><issue>1</issue><spage>147</spage><epage>157</epage><pages>147-157</pages><issn>0092-8674</issn><eissn>1097-4172</eissn><abstract>A cell-free system based on cytosols of normally growing cells is established that reproduces aspects of the apoptotic program in vitro. The apoptotic program is initiated by addition of dATP. Fractionation of cytosol yielded a 15 kDa protein that is required for in vitro apoptosis. The absorption spectrum and protein sequence revealed that this protein is cytochrome c. Elimination of cytochrome c from cytosol by immunodepletion, or inclusion of sucrose to stabilize mitochondria during cytosol preparation, diminished the apoptotic activity. Adding back cytochrome c to the cytochrome c–depleted extracts restored their apoptotic activity. Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8689682</pmid><doi>10.1016/S0092-8674(00)80085-9</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Cell Press Free Archives; Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Adenosine Triphosphate - metabolism Amino Acid Sequence Antibodies, Monoclonal Apoptosis - drug effects Caspase 3 Caspases Cell Extracts - chemistry Cell Extracts - pharmacology Cell-Free System Cysteine Endopeptidases - metabolism Cytochrome c Group - immunology Cytochrome c Group - isolation & purification Cytochrome c Group - metabolism Cytosol - chemistry Cytosol - metabolism DNA - drug effects DNA - metabolism Enzyme Precursors - metabolism HeLa Cells - cytology HeLa Cells - drug effects HeLa Cells - enzymology Humans Molecular Sequence Data |
| title | Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c |
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