Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy
Identifieur interne : 000109 ( Pmc/Corpus ); précédent : 000108; suivant : 000110Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy
Auteurs : Johnson J. Inbaraj ; Thomas B. Cardon ; Mikhail Laryukhin ; Stuart M. Grosser ; Gary A. LoriganSource :
- Journal of the American Chemical Society [ 0002-7863 ] ; 2006.
Abstract
This paper reports on the development of a new structural biology technique for determining the membrane topology of an integral membrane protein inserted into magnetically aligned phospholipid bilayers (bicelles) using EPR spectroscopy. The nitroxide spin probe, 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) was attached to the pore-lining transmembrane domain (M2δ) of the nicotinic acetylcholine receptor (AChR) and incorporated into a bicelle. The corresponding EPR spectra revealed hyperfine splittings that were highly dependent on the macroscopic orientation of the bicelles with respect to the static magnetic field. The helical tilt of the peptide can be easily calculated using the hyperfine splittings gleaned from the orientational dependent EPR spectra. A helical tilt of 14° was calculated for the M2δ peptide with respect to the bilayer normal of the membrane, which agrees well with previous 15N solid-state NMR studies. The helical tilt of the peptide was verified by simulating the corresponding EPR spectra using the standardized MOMD approach. This new method is advantageous because: (1) bicelle samples are easy to prepare, (2) the helical tilt can be directly calculated from the orientational-dependent hyperfine splitting in the EPR spectra, and (3) EPR spectroscopy is approximately 1000 fold more sensitive than 15N solid-state NMR spectroscopy; thus, the helical tilt of an integral membrane peptide can be determined with only 100 μg of peptide. The helical tilt can be determined more accurately by placing TOAC spin labels at several positions with this technique.
Url:
DOI: 10.1021/ja0622204
PubMed: 16848493
PubMed Central: 2533427
Links to Exploration step
PMC:2533427Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy</title>
<author><name sortKey="Inbaraj, Johnson J" sort="Inbaraj, Johnson J" uniqKey="Inbaraj J" first="Johnson J." last="Inbaraj">Johnson J. Inbaraj</name>
</author>
<author><name sortKey="Cardon, Thomas B" sort="Cardon, Thomas B" uniqKey="Cardon T" first="Thomas B." last="Cardon">Thomas B. Cardon</name>
</author>
<author><name sortKey="Laryukhin, Mikhail" sort="Laryukhin, Mikhail" uniqKey="Laryukhin M" first="Mikhail" last="Laryukhin">Mikhail Laryukhin</name>
</author>
<author><name sortKey="Grosser, Stuart M" sort="Grosser, Stuart M" uniqKey="Grosser S" first="Stuart M." last="Grosser">Stuart M. Grosser</name>
</author>
<author><name sortKey="Lorigan, Gary A" sort="Lorigan, Gary A" uniqKey="Lorigan G" first="Gary A." last="Lorigan">Gary A. Lorigan</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">16848493</idno>
<idno type="pmc">2533427</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2533427</idno>
<idno type="RBID">PMC:2533427</idno>
<idno type="doi">10.1021/ja0622204</idno>
<date when="2006">2006</date>
<idno type="wicri:Area/Pmc/Corpus">000109</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000109</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy</title>
<author><name sortKey="Inbaraj, Johnson J" sort="Inbaraj, Johnson J" uniqKey="Inbaraj J" first="Johnson J." last="Inbaraj">Johnson J. Inbaraj</name>
</author>
<author><name sortKey="Cardon, Thomas B" sort="Cardon, Thomas B" uniqKey="Cardon T" first="Thomas B." last="Cardon">Thomas B. Cardon</name>
</author>
<author><name sortKey="Laryukhin, Mikhail" sort="Laryukhin, Mikhail" uniqKey="Laryukhin M" first="Mikhail" last="Laryukhin">Mikhail Laryukhin</name>
</author>
<author><name sortKey="Grosser, Stuart M" sort="Grosser, Stuart M" uniqKey="Grosser S" first="Stuart M." last="Grosser">Stuart M. Grosser</name>
</author>
<author><name sortKey="Lorigan, Gary A" sort="Lorigan, Gary A" uniqKey="Lorigan G" first="Gary A." last="Lorigan">Gary A. Lorigan</name>
</author>
</analytic>
<series><title level="j">Journal of the American Chemical Society</title>
<idno type="ISSN">0002-7863</idno>
<idno type="eISSN">1520-5126</idno>
<imprint><date when="2006">2006</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p id="P1">This paper reports on the development of a new structural biology technique for determining the membrane topology of an integral membrane protein inserted into magnetically aligned phospholipid bilayers (bicelles) using EPR spectroscopy. The nitroxide spin probe, 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) was attached to the pore-lining transmembrane domain (M2δ) of the nicotinic acetylcholine receptor (AChR) and incorporated into a bicelle. The corresponding EPR spectra revealed hyperfine splittings that were highly dependent on the macroscopic orientation of the bicelles with respect to the static magnetic field. The helical tilt of the peptide can be easily calculated using the hyperfine splittings gleaned from the orientational dependent EPR spectra. A helical tilt of 14° was calculated for the M2δ peptide with respect to the bilayer normal of the membrane, which agrees well with previous <sup>15</sup>
N solid-state NMR studies. The helical tilt of the peptide was verified by simulating the corresponding EPR spectra using the standardized MOMD approach. This new method is advantageous because: (1) bicelle samples are easy to prepare, (2) the helical tilt can be directly calculated from the orientational-dependent hyperfine splitting in the EPR spectra, and (3) EPR spectroscopy is approximately 1000 fold more sensitive than <sup>15</sup>
N solid-state NMR spectroscopy; thus, the helical tilt of an integral membrane peptide can be determined with only 100 μg of peptide. The helical tilt can be determined more accurately by placing TOAC spin labels at several positions with this technique.</p>
</div>
</front>
</TEI>
<pmc article-type="research-article" xml:lang="EN"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<pmc-dir>properties manuscript</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-journal-id">7503056</journal-id>
<journal-id journal-id-type="pubmed-jr-id">4435</journal-id>
<journal-id journal-id-type="nlm-ta">J Am Chem Soc</journal-id>
<journal-title>Journal of the American Chemical Society</journal-title>
<issn pub-type="ppub">0002-7863</issn>
<issn pub-type="epub">1520-5126</issn>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">16848493</article-id>
<article-id pub-id-type="pmc">2533427</article-id>
<article-id pub-id-type="doi">10.1021/ja0622204</article-id>
<article-id pub-id-type="manuscript">NIHMS61213</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Inbaraj</surname>
<given-names>Johnson J.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Cardon</surname>
<given-names>Thomas B.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Laryukhin</surname>
<given-names>Mikhail</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Grosser</surname>
<given-names>Stuart M.</given-names>
</name>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Lorigan</surname>
<given-names>Gary A.</given-names>
</name>
</contrib>
<aff id="A1">Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, E-mail:<email>lorigag@muohio.edu</email>
</aff>
</contrib-group>
<pub-date pub-type="nihms-submitted"><day>5</day>
<month>8</month>
<year>2008</year>
</pub-date>
<pub-date pub-type="ppub"><day>26</day>
<month>7</month>
<year>2006</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>11</day>
<month>9</month>
<year>2008</year>
</pub-date>
<volume>128</volume>
<issue>29</issue>
<fpage>9549</fpage>
<lpage>9554</lpage>
<abstract><p id="P1">This paper reports on the development of a new structural biology technique for determining the membrane topology of an integral membrane protein inserted into magnetically aligned phospholipid bilayers (bicelles) using EPR spectroscopy. The nitroxide spin probe, 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) was attached to the pore-lining transmembrane domain (M2δ) of the nicotinic acetylcholine receptor (AChR) and incorporated into a bicelle. The corresponding EPR spectra revealed hyperfine splittings that were highly dependent on the macroscopic orientation of the bicelles with respect to the static magnetic field. The helical tilt of the peptide can be easily calculated using the hyperfine splittings gleaned from the orientational dependent EPR spectra. A helical tilt of 14° was calculated for the M2δ peptide with respect to the bilayer normal of the membrane, which agrees well with previous <sup>15</sup>
N solid-state NMR studies. The helical tilt of the peptide was verified by simulating the corresponding EPR spectra using the standardized MOMD approach. This new method is advantageous because: (1) bicelle samples are easy to prepare, (2) the helical tilt can be directly calculated from the orientational-dependent hyperfine splitting in the EPR spectra, and (3) EPR spectroscopy is approximately 1000 fold more sensitive than <sup>15</sup>
N solid-state NMR spectroscopy; thus, the helical tilt of an integral membrane peptide can be determined with only 100 μg of peptide. The helical tilt can be determined more accurately by placing TOAC spin labels at several positions with this technique.</p>
</abstract>
<contract-num rid="GM1">R01 GM080542-01A2</contract-num>
<contract-sponsor id="GM1">National Institute of General Medical Sciences : NIGMS</contract-sponsor>
</article-meta>
</front>
</pmc>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Terre/explor/ThuliumV1/Data/Pmc/Corpus
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000109 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Corpus/biblio.hfd -nk 000109 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Wicri/Terre |area= ThuliumV1 |flux= Pmc |étape= Corpus |type= RBID |clé= PMC:2533427 |texte= Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Corpus/RBID.i -Sk "pubmed:16848493" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Corpus/biblio.hfd \ | NlmPubMed2Wicri -a ThuliumV1
![]() | This area was generated with Dilib version V0.6.21. | ![]() |