Addgene - Revision history

Bradley Monk at 03:20, 10 May 2015

2015-05-10T03:20:27Z

← Older revision		Revision as of 20:20, 9 May 2015
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	==Dronpa==		==Dronpa==

	{{Template:Addgene\|Dronpa-GluR2\|62735\|~~[[File:Dronpa_GluR2~~.png~~\|400px]]~~\|Mark Mayford}}		{{Template:Addgene\|Dronpa-GluR2\|62735\|file=Dronpa GluR2.png\|Mark Mayford}}

	;: [http://www.addgene.org/62735/ addgene: Dronpa-GluR2]		;: [http://www.addgene.org/62735/ addgene: Dronpa-GluR2]

Bradley Monk at 03:11, 10 May 2015

2015-05-10T03:11:31Z

← Older revision		Revision as of 20:11, 9 May 2015
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	==Dronpa==		==Dronpa==

			{{Template:Addgene\|Dronpa-GluR2\|62735\|[[File:Dronpa_GluR2.png\|400px]]\|Mark Mayford}}

	;: [http://www.addgene.org/62735/ addgene: Dronpa-GluR2]		;: [http://www.addgene.org/62735/ addgene: Dronpa-GluR2]

Bradley Monk at 02:33, 10 May 2015

2015-05-10T02:33:36Z

← Older revision		Revision as of 19:33, 9 May 2015
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			==pTRE_GFP-GluR1==


	==Dronpa==		==Dronpa==

	[http://www.addgene.org/62735/ addgene: Dronpa-GluR2]		;: [http://www.addgene.org/62735/ addgene: Dronpa-GluR2]

	[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]		[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]

Bradley Monk at 02:32, 10 May 2015

2015-05-10T02:32:46Z

← Older revision		Revision as of 19:32, 9 May 2015
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	==Dronpa==		==Dronpa==

	[[http://www.addgene.org/62735/ addgene - ~~Dronpa]~~]		[http://www.addgene.org/62735/ addgene: Dronpa-GluR2]

	[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]		[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]

Bradley Monk at 02:32, 10 May 2015

2015-05-10T02:32:18Z

← Older revision		Revision as of 19:32, 9 May 2015
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	==Dronpa==		==Dronpa==

			[[http://www.addgene.org/62735/ addgene - Dronpa]]

	[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]		[[File:Dronpa structure animation.gif\|thumb\|width=251\|\|An animation of the structure of the dark state of dronpa protein]]
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	When a weakly tetrameric form of Dronpa photoswitches, its oligomerization affinity changes. This was used to enable optical control over the activity of enzymes. Specifically, two Dronpa domains attached to the ends of a protein oligomerized and blocked enzyme activity in the dark, but monomerized after illumination, enabling enzyme activity.<ref name=Zhou2012/>		When a weakly tetrameric form of Dronpa photoswitches, its oligomerization affinity changes. This was used to enable optical control over the activity of enzymes. Specifically, two Dronpa domains attached to the ends of a protein oligomerized and blocked enzyme activity in the dark, but monomerized after illumination, enabling enzyme activity.<ref name=Zhou2012/>

	====References====		==References==

	{{reflist}}		{{reflist}}

	[[Category:Proteins]]		[[Category:Proteins]]

Bradley Monk: Created page with " ==Dronpa== An animation of the structure of the dark state of dronpa protein '''Dronpa''' is a reversibly switc..."

2015-05-10T00:46:32Z

Created page with " ==Dronpa== thumb|width=251||An animation of the structure of the dark state of dronpa protein '''Dronpa''' is a reversibly switc..."

New page

==Dronpa==

[[File:Dronpa structure animation.gif|thumb|width=251||An animation of the structure of the dark state of dronpa protein]]
'''Dronpa''' is a reversibly switchable [[photoactivatable fluorescent protein]] that is 2.5 times as bright as [[EGFP]].<ref name=Day2010>{{cite doi|10.1039/b901966a}}</ref><ref name=Ando2004>{{cite doi|10.1126/science.1102506}}</ref> Dronpa gets switched off by strong illumination with 488 nm (blue) light and this can be reversed by weak 405 nm UV light.<ref name=Day2010/> A single dronpa molecule can switched on and off over 100 times.<ref name=Habuchi2005>{{cite doi|10.1073/pnas.0500489102}}</ref> It has an excitation peak at 503 nm and an emission peak at 518 nm.

A tetrameric,<ref name=Zhou2012>{{cite doi|10.1126/science.1226854}}</ref> reversibly switchable fluorescent protein was discovered in a [[cDNA]] screen of a stony coral ([[Pectiniidae]]). A monomeric variant of this protein was named "Dronpa" after "''Dron''" a ninja term for vanishing and ''pa'' for photoactivation.<ref name=Ando2004/>

[[File:Dronpa chromophore states.png|thumb|On-cis (green) and Off-trans (yellow) states of the chromophore. Nearby residues that move are also shown. Clockwise from the top: Arg66, Val157, Ser142, Cys-Trp-Gly chromophore.]]
Dronpa is 257 amino acids long and is a 28.8 kDa monomer. Dronpa is 76% similar in sequence to GFP<ref name=Ando2004/> and shares a similar structure with an 11 stranded β-barrel (a β-can) enclosing an α-helix.<ref name=Andreson2007>{{cite doi|10.1073/pnas.0700629104 }}</ref> The chromophore is formed autocatalytically from residues Cys62,Tyr63 and Gly64.<ref name=Andreson2007/><ref name=Stiel2007>{{cite doi|10.1042/BJ20061401}}</ref> The on state of the dronpa molecule has the chromophore in a cis conformation while the off state chromophore exists in the trans conformation. Several other residues in the vicinity of the chromophore also move during the on-off transition resulting a very different electrostatic environment.<ref name=Andreson2007/>

====Applications====
Dronpa's fast dynamics and stability under repeated cycles of switching make it one of the more important switchable fluorescent proteins.<ref name=Day2010/> It is used in [[super resolution microscopy]] techniques like [[Super resolution microscopy#Photoactivated localization microscopy (PALM) and Fluorescence Photoactivation Localization Microscopy (fPALM)|PALM/STORM]]. It can also be used to track fast dynamics of proteins in cells.

When a weakly tetrameric form of Dronpa photoswitches, its oligomerization affinity changes. This was used to enable optical control over the activity of enzymes. Specifically, two Dronpa domains attached to the ends of a protein oligomerized and blocked enzyme activity in the dark, but monomerized after illumination, enabling enzyme activity.<ref name=Zhou2012/>

====References====

{{reflist}}

[[Category:Proteins]]