MALDI Imaging and in Situ Identification of Integral Membrane Proteins from Rat Brain Tissue Sections
Transmembrane proteins are greatly underrepresented in data generated by imaging mass spectrometry (IMS) because of analytical challenges related to their size and solubility. Here, we present the first example of MALDI IMS of two highly modified multitransmembrane domain proteins, myelin proteolipi...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2013-08, Vol.85 (15), p.7191-7196 |
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| creator | Nicklay, Joshua J Harris, Glenn A Schey, Kevin L Caprioli, Richard M |
| description | Transmembrane proteins are greatly underrepresented in data generated by imaging mass spectrometry (IMS) because of analytical challenges related to their size and solubility. Here, we present the first example of MALDI IMS of two highly modified multitransmembrane domain proteins, myelin proteolipid protein (PLP, 30 kDa) and DM-20 (26 kDa), from various regions of rat brain, namely, the cerebrum, cerebellum, and medulla. We utilize a novel tissue pretreatment aimed at transmembrane protein enrichment to show the in situ distribution of fatty acylation of these proteins, particularly of post-translational palmitoylation. Additionally, we demonstrate the utility of protease-encapsulated hydrogels for spatially localized on-tissue protein digestion and peptide extraction for subsequent direct coupling to LC-MS/MS for protein identification. |
| doi_str_mv | 10.1021/ac400902h |
| format | Article |
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Here, we present the first example of MALDI IMS of two highly modified multitransmembrane domain proteins, myelin proteolipid protein (PLP, 30 kDa) and DM-20 (26 kDa), from various regions of rat brain, namely, the cerebrum, cerebellum, and medulla. We utilize a novel tissue pretreatment aimed at transmembrane protein enrichment to show the in situ distribution of fatty acylation of these proteins, particularly of post-translational palmitoylation. 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Chem</addtitle><description>Transmembrane proteins are greatly underrepresented in data generated by imaging mass spectrometry (IMS) because of analytical challenges related to their size and solubility. Here, we present the first example of MALDI IMS of two highly modified multitransmembrane domain proteins, myelin proteolipid protein (PLP, 30 kDa) and DM-20 (26 kDa), from various regions of rat brain, namely, the cerebrum, cerebellum, and medulla. We utilize a novel tissue pretreatment aimed at transmembrane protein enrichment to show the in situ distribution of fatty acylation of these proteins, particularly of post-translational palmitoylation. Additionally, we demonstrate the utility of protease-encapsulated hydrogels for spatially localized on-tissue protein digestion and peptide extraction for subsequent direct coupling to LC-MS/MS for protein identification.</description><subject>acylation</subject><subject>Animals</subject><subject>Brain</subject><subject>Brain - metabolism</subject><subject>Cerebellum</subject><subject>cerebrum</subject><subject>Digestion</subject><subject>Enrichment</subject><subject>hydrocolloids</subject><subject>image analysis</subject><subject>Imaging</subject><subject>Lipids</subject><subject>Mass spectrometry</subject><subject>Molecular Imaging - methods</subject><subject>Myelin</subject><subject>Myelin Proteolipid Protein - metabolism</subject><subject>myelin sheath</subject><subject>palmitoylation</subject><subject>Peptides</subject><subject>Proteins</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Rodents</subject><subject>solubility</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</subject><subject>Tissues</subject><subject>transmembrane proteins</subject><subject>Trypsin - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkk9v1DAQxS0EokvhwBdAlhBSOQTGduLEF6RS_kXaCkTLOXKcydbVxi52Uolvz1RbVgUOPfkwP795b_QYey7gjQAp3lpXAhiQFw_YSlQSCt008iFbAYAqZA1wwJ7kfAkgBAj9mB1I1UgjTbVieHq8_tDydrIbHzbchoH7wM_8vPB2wDD70Ts7-xh4HHkbZtwku-WnOPXJBuTfUpzRh8zHFCf-3c78fbIkcO5zXpCfobv5m5-yR6PdZnx2-x6yH58-np98KdZfP7cnx-vClsbMxSiUNMKSNYASBugdWlRWGxxR164WNWqrxTD2zlBCJyvK0xtnQfU0keqQvdvpXi39hIOjAGS3u0p-sulXF63v_p4Ef9Ft4nWn6kZCU5LA0a1Aij8XzHM3-exwu6WwcckdGQNZNXUj7kWFNlKpSpdwP1qKRumSHBD68h_0Mi4p0NGIkqAqSeuJer2jXIo5Jxz3EQV0N5Xo9pUg9sXdm-zJPx0g4NUOsC7f2faf0G9O6rus</recordid><startdate>20130806</startdate><enddate>20130806</enddate><creator>Nicklay, Joshua J</creator><creator>Harris, Glenn A</creator><creator>Schey, Kevin L</creator><creator>Caprioli, Richard M</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20130806</creationdate><title>MALDI Imaging and in Situ Identification of Integral Membrane Proteins from Rat Brain Tissue Sections</title><author>Nicklay, Joshua J ; Harris, Glenn A ; Schey, Kevin L ; Caprioli, Richard M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a499t-f13291a2920040d0bceae3a69efe67c717e6a61dfbc9882c25101b9ca03be6a23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>acylation</topic><topic>Animals</topic><topic>Brain</topic><topic>Brain - metabolism</topic><topic>Cerebellum</topic><topic>cerebrum</topic><topic>Digestion</topic><topic>Enrichment</topic><topic>hydrocolloids</topic><topic>image analysis</topic><topic>Imaging</topic><topic>Lipids</topic><topic>Mass spectrometry</topic><topic>Molecular Imaging - methods</topic><topic>Myelin</topic><topic>Myelin Proteolipid Protein - metabolism</topic><topic>myelin sheath</topic><topic>palmitoylation</topic><topic>Peptides</topic><topic>Proteins</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Rodents</topic><topic>solubility</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</topic><topic>Tissues</topic><topic>transmembrane proteins</topic><topic>Trypsin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nicklay, Joshua J</creatorcontrib><creatorcontrib>Harris, Glenn A</creatorcontrib><creatorcontrib>Schey, Kevin L</creatorcontrib><creatorcontrib>Caprioli, Richard M</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nicklay, Joshua J</au><au>Harris, Glenn A</au><au>Schey, Kevin L</au><au>Caprioli, Richard M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MALDI Imaging and in Situ Identification of Integral Membrane Proteins from Rat Brain Tissue Sections</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. 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| source | MEDLINE; ACS Publications |
| subjects | acylation Animals Brain Brain - metabolism Cerebellum cerebrum Digestion Enrichment hydrocolloids image analysis Imaging Lipids Mass spectrometry Molecular Imaging - methods Myelin Myelin Proteolipid Protein - metabolism myelin sheath palmitoylation Peptides Proteins Rats Rats, Sprague-Dawley Rodents solubility Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods Tissues transmembrane proteins Trypsin - metabolism |
| title | MALDI Imaging and in Situ Identification of Integral Membrane Proteins from Rat Brain Tissue Sections |
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