Transition metal ion FRET to measure short-range distances at the intracellular surface of the plasma membrane
Identifieur interne : 000065 ( Pmc/Curation ); précédent : 000064; suivant : 000066Transition metal ion FRET to measure short-range distances at the intracellular surface of the plasma membrane
Auteurs : Sharona E. Gordon ; Eric N. Senning ; Teresa K. Aman ; William N. ZagottaSource :
- The Journal of General Physiology [ 0022-1295 ] ; 2016.
Abstract
A novel method is presented to measure short distances in cell plasma membranes using transition metal ion FRET with metal ions bound to introduced sites in the membrane.
Url:
DOI: 10.1085/jgp.201511530
PubMed: 26755772
PubMed Central: 4727948
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Transition metal ion FRET to measure short-range distances at the intracellular surface of the plasma membrane</title>
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<front><div type="abstract" xml:lang="en"><p>A novel method is presented to measure short distances in cell plasma membranes using transition metal ion FRET with metal ions bound to introduced sites in the membrane.</p>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Gen Physiol</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Gen. Physiol</journal-id>
<journal-id journal-id-type="hwp">jgp</journal-id>
<journal-id journal-id-type="publisher-id">jgp</journal-id>
<journal-title-group><journal-title>The Journal of General Physiology</journal-title>
</journal-title-group>
<issn pub-type="ppub">0022-1295</issn>
<issn pub-type="epub">1540-7748</issn>
<publisher><publisher-name>The Rockefeller University Press</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">26755772</article-id>
<article-id pub-id-type="pmc">4727948</article-id>
<article-id pub-id-type="publisher-id">201511530</article-id>
<article-id pub-id-type="doi">10.1085/jgp.201511530</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Methods and Approaches</subject>
</subj-group>
</article-categories>
<title-group><article-title>Transition metal ion FRET to measure short-range distances at the intracellular surface of the plasma membrane</article-title>
<alt-title alt-title-type="short">Measuring intramembrane distances with tmFRET</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author" corresp="yes"><name><surname>Gordon</surname>
<given-names>Sharona E.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Senning</surname>
<given-names>Eric N.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Aman</surname>
<given-names>Teresa K.</given-names>
</name>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Zagotta</surname>
<given-names>William N.</given-names>
</name>
</contrib>
</contrib-group>
<aff><institution>Department of Physiology and Biophysics, University of Washington, Seattle, WA 98195</institution>
</aff>
<author-notes><corresp>Correspondence to Sharona E. Gordon: <email>seg@uw.edu</email>
; or William N. Zagotta: <email>zagotta@uw.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>2</month>
<year>2016</year>
</pub-date>
<volume>147</volume>
<issue>2</issue>
<fpage>189</fpage>
<lpage>200</lpage>
<history><date date-type="received"><day>14</day>
<month>10</month>
<year>2015</year>
</date>
<date date-type="accepted"><day>21</day>
<month>12</month>
<year>2015</year>
</date>
</history>
<permissions><copyright-statement>© 2016 Gordon et al.</copyright-statement>
<copyright-year>2016</copyright-year>
<license license-type="openaccess"><license-p>This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see <ext-link ext-link-type="uri" xlink:href="http://www.rupress.org/terms">http://www.rupress.org/terms</ext-link>
). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc-sa/3.0/">http://creativecommons.org/licenses/by-nc-sa/3.0/</ext-link>
).</license-p>
</license>
</permissions>
<self-uri xlink:role="icon" xlink:href="JGP_201511530_thumb.gif"></self-uri>
<abstract abstract-type="precis"><p>A novel method is presented to measure short distances in cell plasma membranes using transition metal ion FRET with metal ions bound to introduced sites in the membrane.</p>
</abstract>
<abstract><p>Biological membranes are complex assemblies of lipids and proteins that serve as platforms for cell signaling. We have developed a novel method for measuring the structure and dynamics of the membrane based on fluorescence resonance energy transfer (FRET). The method marries four technologies: (1) unroofing cells to isolate and access the cytoplasmic leaflet of the plasma membrane; (2) patch-clamp fluorometry (PCF) to measure currents and fluorescence simultaneously from a membrane patch; (3) a synthetic lipid with a metal-chelating head group to decorate the membrane with metal-binding sites; and (4) transition metal ion FRET (tmFRET) to measure short distances between a fluorescent probe and a transition metal ion on the membrane. We applied this method to measure the density and affinity of native and introduced metal-binding sites in the membrane. These experiments pave the way for measuring structural rearrangements of membrane proteins relative to the membrane.</p>
</abstract>
</article-meta>
</front>
</pmc>
</record>
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