Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane

Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromopho...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Yuka eJinno [verfasserIn] Keiko eShoda [verfasserIn] Emiliano eRial-Verde [verfasserIn] Rafael eYuste [verfasserIn] Miyawaki eAtsushi [verfasserIn] Hidekazu eTsutsui [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2014

Schlagwörter:	Mutagenesis electrostatic potential second harmonic generation fluoresence protein Kras4B

Übergeordnetes Werk:	In: Frontiers in Molecular Neuroscience - Frontiers Media S.A., 2008, 7(2014)
Übergeordnetes Werk:	volume:7 ; year:2014

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fnmol.2014.00093

Katalog-ID:	DOAJ02541240X

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520			\|a Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the membrane-cytoplasm interface using the fluorescent protein mVenus. After adding a farnesylation motif to the C-terminus of mVenus, nine amino acids composing its -barrel surface were replaced by lysine, forming an electrostatic patch. This protein (mVe9Knus-CVIM) was efficiently targeted to the plasma membrane in a geometrically defined manner and exhibited SHG in HEK293 cells. In agreement with its design, mVe9Knus-CVIM hyperpolarizability was oriented at a small angle (~7.3º) from the membrane normal. Genetically-encoded SHG chromophores could serve as a molecular platform for imaging membrane potential.
650		4	\|a Mutagenesis
650		4	\|a electrostatic potential
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language	English
source	In Frontiers in Molecular Neuroscience 7(2014) volume:7 year:2014
sourceStr	In Frontiers in Molecular Neuroscience 7(2014) volume:7 year:2014
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Mutagenesis electrostatic potential second harmonic generation fluoresence protein Kras4B Neurosciences. Biological psychiatry. Neuropsychiatry
isfreeaccess_bool	true
container_title	Frontiers in Molecular Neuroscience
authorswithroles_txt_mv	Yuka eJinno @@aut@@ Keiko eShoda @@aut@@ Emiliano eRial-Verde @@aut@@ Rafael eYuste @@aut@@ Miyawaki eAtsushi @@aut@@ Hidekazu eTsutsui @@aut@@
publishDateDaySort_date	2014-01-01T00:00:00Z
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callnumber-first	R - Medicine
author	Yuka eJinno
spellingShingle	Yuka eJinno misc RC321-571 misc Mutagenesis misc electrostatic potential misc second harmonic generation misc fluoresence protein misc Kras4B misc Neurosciences. Biological psychiatry. Neuropsychiatry Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane
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topic_title	RC321-571 Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane Mutagenesis electrostatic potential second harmonic generation fluoresence protein Kras4B
topic	misc RC321-571 misc Mutagenesis misc electrostatic potential misc second harmonic generation misc fluoresence protein misc Kras4B misc Neurosciences. Biological psychiatry. Neuropsychiatry
topic_unstemmed	misc RC321-571 misc Mutagenesis misc electrostatic potential misc second harmonic generation misc fluoresence protein misc Kras4B misc Neurosciences. Biological psychiatry. Neuropsychiatry
topic_browse	misc RC321-571 misc Mutagenesis misc electrostatic potential misc second harmonic generation misc fluoresence protein misc Kras4B misc Neurosciences. Biological psychiatry. Neuropsychiatry
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title	Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane
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title_full	Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane
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title_sort	engineering a genetically-encoded shg chromophore by electrostatic targeting to the membrane
callnumber	RC321-571
title_auth	Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane
abstract	Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the membrane-cytoplasm interface using the fluorescent protein mVenus. After adding a farnesylation motif to the C-terminus of mVenus, nine amino acids composing its -barrel surface were replaced by lysine, forming an electrostatic patch. This protein (mVe9Knus-CVIM) was efficiently targeted to the plasma membrane in a geometrically defined manner and exhibited SHG in HEK293 cells. In agreement with its design, mVe9Knus-CVIM hyperpolarizability was oriented at a small angle (~7.3º) from the membrane normal. Genetically-encoded SHG chromophores could serve as a molecular platform for imaging membrane potential.
abstractGer	Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the membrane-cytoplasm interface using the fluorescent protein mVenus. After adding a farnesylation motif to the C-terminus of mVenus, nine amino acids composing its -barrel surface were replaced by lysine, forming an electrostatic patch. This protein (mVe9Knus-CVIM) was efficiently targeted to the plasma membrane in a geometrically defined manner and exhibited SHG in HEK293 cells. In agreement with its design, mVe9Knus-CVIM hyperpolarizability was oriented at a small angle (~7.3º) from the membrane normal. Genetically-encoded SHG chromophores could serve as a molecular platform for imaging membrane potential.
abstract_unstemmed	Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the membrane-cytoplasm interface using the fluorescent protein mVenus. After adding a farnesylation motif to the C-terminus of mVenus, nine amino acids composing its -barrel surface were replaced by lysine, forming an electrostatic patch. This protein (mVe9Knus-CVIM) was efficiently targeted to the plasma membrane in a geometrically defined manner and exhibited SHG in HEK293 cells. In agreement with its design, mVe9Knus-CVIM hyperpolarizability was oriented at a small angle (~7.3º) from the membrane normal. Genetically-encoded SHG chromophores could serve as a molecular platform for imaging membrane potential.
collection_details	GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700
title_short	Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane
url	https://doi.org/10.3389/fnmol.2014.00093 https://doaj.org/article/bebbce596950480ab5c13ff7f70006d7 http://journal.frontiersin.org/Journal/10.3389/fnmol.2014.00093/full https://doaj.org/toc/1662-5099
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Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane

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