Optimization of Neuropeptide Extraction from the Mouse Hypothalamus
Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments f...
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| Veröffentlicht in: | Journal of proteome research 2007-12, Vol.6 (12), p.4667-4676 |
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| container_issue | 12 |
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| container_title | Journal of proteome research |
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| creator | Che, Fa-Yun Zhang, Xin Berezniuk, Iryna Callaway, Myrasol Lim, Jihyeon Fricker, Lloyd D |
| description | Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70 °C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides. |
| doi_str_mv | 10.1021/pr060690r |
| format | Article |
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We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70 °C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides.</description><identifier>ISSN: 1535-3893</identifier><identifier>EISSN: 1535-3907</identifier><identifier>DOI: 10.1021/pr060690r</identifier><identifier>PMID: 17979226</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Amino Acid Sequence ; Animals ; Chromatography, Liquid ; Female ; Hypothalamus - chemistry ; Hypothalamus - metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Neuropeptides - isolation & purification ; Neuropeptides - metabolism ; Tandem Mass Spectrometry</subject><ispartof>Journal of proteome research, 2007-12, Vol.6 (12), p.4667-4676</ispartof><rights>Copyright © 2007 American Chemical Society</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a313t-22d5cc407dafb599db3181e799c6bb5d254f27f8d117251e974c4dff9189c3bf3</citedby><cites>FETCH-LOGICAL-a313t-22d5cc407dafb599db3181e799c6bb5d254f27f8d117251e974c4dff9189c3bf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/pr060690r$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/pr060690r$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,776,780,2752,27053,27901,27902,56713,56763</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17979226$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Che, Fa-Yun</creatorcontrib><creatorcontrib>Zhang, Xin</creatorcontrib><creatorcontrib>Berezniuk, Iryna</creatorcontrib><creatorcontrib>Callaway, Myrasol</creatorcontrib><creatorcontrib>Lim, Jihyeon</creatorcontrib><creatorcontrib>Fricker, Lloyd D</creatorcontrib><title>Optimization of Neuropeptide Extraction from the Mouse Hypothalamus</title><title>Journal of proteome research</title><addtitle>J. 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The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Chromatography, Liquid</subject><subject>Female</subject><subject>Hypothalamus - chemistry</subject><subject>Hypothalamus - metabolism</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Molecular Sequence Data</subject><subject>Neuropeptides - isolation & purification</subject><subject>Neuropeptides - metabolism</subject><subject>Tandem Mass Spectrometry</subject><issn>1535-3893</issn><issn>1535-3907</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkD1PwzAQhi0EoqUw8AdQFpAYAv6I43hEVaFIhS4wW44_1FRJHWxHovx6DC2wcMud7Eev7h4AzhG8QRCj297DEpYc-gMwRpTQnHDIDn_mipMROAlhDSGiDJJjMEKMM45xOQbTZR-brvmQsXGbzNns2Qze9Sa9apPN3qOX6vvLetdlcWWyJzcEk823vYsr2cpuCKfgyMo2mLN9n4DX-9nLdJ4vlg-P07tFLgkiMcdYU6UKyLS0NeVc1wRVyDDOVVnXVGNaWMxspRFimCLDWaEKbS1HFVektmQCrna5vXdvgwlRdE1Qpm3lxqSlRDJQVKkSeL0DlXcheGNF75tO-q1AUHwZE7_GEnuxDx3qzug_cq8oAZc7QKog1m7wm3TjP0GfGzly-g</recordid><startdate>20071201</startdate><enddate>20071201</enddate><creator>Che, Fa-Yun</creator><creator>Zhang, Xin</creator><creator>Berezniuk, Iryna</creator><creator>Callaway, Myrasol</creator><creator>Lim, Jihyeon</creator><creator>Fricker, Lloyd D</creator><general>American Chemical Society</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>20071201</creationdate><title>Optimization of Neuropeptide Extraction from the Mouse Hypothalamus</title><author>Che, Fa-Yun ; Zhang, Xin ; Berezniuk, Iryna ; Callaway, Myrasol ; Lim, Jihyeon ; Fricker, Lloyd D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a313t-22d5cc407dafb599db3181e799c6bb5d254f27f8d117251e974c4dff9189c3bf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Chromatography, Liquid</topic><topic>Female</topic><topic>Hypothalamus - chemistry</topic><topic>Hypothalamus - metabolism</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Molecular Sequence Data</topic><topic>Neuropeptides - isolation & purification</topic><topic>Neuropeptides - metabolism</topic><topic>Tandem Mass Spectrometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Che, Fa-Yun</creatorcontrib><creatorcontrib>Zhang, Xin</creatorcontrib><creatorcontrib>Berezniuk, Iryna</creatorcontrib><creatorcontrib>Callaway, Myrasol</creatorcontrib><creatorcontrib>Lim, Jihyeon</creatorcontrib><creatorcontrib>Fricker, Lloyd D</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>Journal of proteome research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Che, Fa-Yun</au><au>Zhang, Xin</au><au>Berezniuk, Iryna</au><au>Callaway, Myrasol</au><au>Lim, Jihyeon</au><au>Fricker, Lloyd D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Optimization of Neuropeptide Extraction from the Mouse Hypothalamus</atitle><jtitle>Journal of proteome research</jtitle><addtitle>J. Proteome Res</addtitle><date>2007-12-01</date><risdate>2007</risdate><volume>6</volume><issue>12</issue><spage>4667</spage><epage>4676</epage><pages>4667-4676</pages><issn>1535-3893</issn><eissn>1535-3907</eissn><abstract>Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70 °C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. 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| ispartof | Journal of proteome research, 2007-12, Vol.6 (12), p.4667-4676 |
| issn | 1535-3893 1535-3907 |
| language | eng |
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| source | MEDLINE; American Chemical Society Publications |
| subjects | Amino Acid Sequence Animals Chromatography, Liquid Female Hypothalamus - chemistry Hypothalamus - metabolism Male Mice Mice, Inbred C57BL Molecular Sequence Data Neuropeptides - isolation & purification Neuropeptides - metabolism Tandem Mass Spectrometry |
| title | Optimization of Neuropeptide Extraction from the Mouse Hypothalamus |
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