PMID:10518952 - Revision history

imported>Wikientrybot: change PMID template to TableEdit version

2013-09-15T04:11:01Z

change PMID template to TableEdit version

← Older revision		Revision as of 04:11, 15 September 2013
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	!align=left \|Citation		!align=left \|Citation
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	'''~~No author listed~~''' (1999) Functional reconstitution and characterization of AqpZ, the E. coli water channel protein. ''J. Mol. Biol.'' '''291''':1169-79		'''Borgnia, MJ, Kozono, D, Calamita, G, Maloney, PC and Agre, P''' (1999) Functional reconstitution and characterization of AqpZ, the E. coli water channel protein. ''J. Mol. Biol.'' '''291''':1169-79
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	!align=left \|Abstract		!align=left \|Abstract

imported>Wikientrybot: replace GO_table_reference with annotationlinks

2013-09-01T17:48:18Z

replace GO_table_reference with annotationlinks

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 ==Annotations {{HelpIcon|Help:Literature}}==
-<!--box uid=63ce1649aab8b3180235b0af7618f667.0.X47ab75b61ce88-->
+<annotationlinks/>
 ==EcoliWiki Links {{HelpIcon|Help:Literature}} ==

imported>Wikientrybot: change PMID template to TableEdit version

2013-09-01T17:48:18Z

change PMID template to TableEdit version

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 {{RightTOC}}
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-|{{TableHeadingColor}} |'''Abstract''' || Understanding the selectivity of aquaporin water channels will require structural and functional studies of wild-type and modified proteins; however, expression systems have not previously yielded aquaporins in the necessary milligram quantities. Here we report expression of a histidine-tagged form of Escherichia coli aquaporin-Z (AqpZ) in its homologous expression system. 10-His-AqpZ is solubilized and purified to near homogeneity in a single step with a final yield of approximately 2.5 mg/l of culture. The histidine tag is removed by trypsin, yielding the native protein with the addition of three N-terminal residues, as confirmed by microsequencing. Sucrose gradient sedimentation analysis showed that the native, solubilized AqpZ protein is a trypsin-resistant tetramer. Unlike other known aquaporins, AqpZ tetramers are not readily dissociated by 1% SDS at neutral pH. Hydrophilic reducing agents have a limited effect on the stability of the tetramer in 1% SDS, whereas incubations for more than 24 hours, pH values below 5.6, or exposure to the hydrophobic reducing agent ethanedithiol cause dissociation into monomers. Cys20, but not Cys9, is necessary for the stability of the AqpZ tetramer in SDS. Upon reconstitution into proteoliposomes, AqpZ displays very high osmotic water permeability (pf > or = 10 x 10(-14) cm3 s-1 subunit-1) and low Arrhenius activation energy (Ea = 3.7 kcal/mol), similar to mammalian aquaporin-1 (AQP1). No permeation by glycerol, urea or sorbitol was detected. Expression of native and modified AqpZ in milligram quantities has permitted biophysical characterization of this remarkably stable aquaporin tetramer, which is being utilized for high-resolution structural studies.
+!align=left  |Citation
-</noinclude>
+||
 |-
-|{{TableHeadingColor}} nowrap |'''Links''' ||  [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10518952 PubMed]
+!align=left  |Keywords
 |}
+<!--box uid=d41d8cd98f00b204e9800998ecf8427e.131884.I52237de1edd7e-->
 ==Significance {{HelpIcon|Help:Literature}} ==
 {{LitSignificance}}

127.0.0.1: /Volumes/r3/t3/ul/working/touchTables/touch_all_tables.php

2010-06-17T19:45:53Z

/Volumes/r3/t3/ul/working/touchTables/touch_all_tables.php

← Older revision		Revision as of 19:45, 17 June 2010
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	{\| id="X47ab75b61ce88" class=" tableEdit GO_table_reference"		{\| id="X47ab75b61ce88" class=" tableEdit GO_table_reference"
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	!\|Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status		!\|Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status

127.0.0.1: Update TableEdit tables.

2010-01-09T03:46:02Z

Update TableEdit tables.

← Older revision		Revision as of 03:46, 9 January 2010
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	!\|Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status		!\|Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status

	\|-class=~~'sortbottom'~~		\|- class="tableEdit_footer"
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imported>Wikientrybot: Fix links due to TableEditLinks.php changes.

2009-02-19T23:00:48Z

Fix links due to TableEditLinks.php changes.

← Older revision		Revision as of 23:00, 19 February 2009
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	<!--		<!--
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	\|- {{table heading style}}		\|- {{table heading style}}
	!Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status		!\|Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status

	\|-class='sortbottom'		\|-class='sortbottom'
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127.0.0.1: Edited by wikibot

2008-04-30T13:08:02Z

Edited by wikibot

← Older revision		Revision as of 13:08, 30 April 2008
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	!Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status		!Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status
	\|-class='sortbottom'		\|-class='sortbottom'
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127.0.0.1: Edited by wikibot

2008-04-25T22:46:45Z

Edited by wikibot

← Older revision		Revision as of 22:46, 25 April 2008
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	\|{{TableHeadingColor}} nowrap \|'''Links''' \|\| [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10518952 PubMed]		\|{{TableHeadingColor}} nowrap \|'''Links''' \|\| [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10518952 PubMed]
	\|}		\|}
	~~</protect>~~
	==Significance {{HelpIcon\|Help:Literature}} ==		==Significance {{HelpIcon\|Help:Literature}} ==
	{{LitSignificance}}		{{LitSignificance}}
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	==Annotations {{HelpIcon\|Help:Literature}}==		==Annotations {{HelpIcon\|Help:Literature}}==
	~~<protect>~~<!--box uid=63ce1649aab8b3180235b0af7618f667.0.X47ab75b61ce88-->		<!--box uid=63ce1649aab8b3180235b0af7618f667.0.X47ab75b61ce88-->
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	==EcoliWiki Links {{HelpIcon\|Help:Literature}} ==		==EcoliWiki Links {{HelpIcon\|Help:Literature}} ==

165.91.51.61: New PMID: Page!

2008-02-07T21:18:46Z

New PMID: Page!

New page

<protect>
{{RightTOC}}

<span style = 'font-size: large'> '''Borgnia, MJ, Kozono, D, Calamita, G, Maloney, PC and Agre, P''' (1999) Functional reconstitution and characterization of AqpZ, the E. coli water channel protein.''J. Mol. Biol.'' '''291''':1169-79</span>

{|{{Prettytable}} align = 'center' width= '70%'
|-
|{{TableHeadingColor}} |'''Abstract''' || Understanding the selectivity of aquaporin water channels will require structural and functional studies of wild-type and modified proteins; however, expression systems have not previously yielded aquaporins in the necessary milligram quantities. Here we report expression of a histidine-tagged form of Escherichia coli aquaporin-Z (AqpZ) in its homologous expression system. 10-His-AqpZ is solubilized and purified to near homogeneity in a single step with a final yield of approximately 2.5 mg/l of culture. The histidine tag is removed by trypsin, yielding the native protein with the addition of three N-terminal residues, as confirmed by microsequencing. Sucrose gradient sedimentation analysis showed that the native, solubilized AqpZ protein is a trypsin-resistant tetramer. Unlike other known aquaporins, AqpZ tetramers are not readily dissociated by 1% SDS at neutral pH. Hydrophilic reducing agents have a limited effect on the stability of the tetramer in 1% SDS, whereas incubations for more than 24 hours, pH values below 5.6, or exposure to the hydrophobic reducing agent ethanedithiol cause dissociation into monomers. Cys20, but not Cys9, is necessary for the stability of the AqpZ tetramer in SDS. Upon reconstitution into proteoliposomes, AqpZ displays very high osmotic water permeability (pf > or = 10 x 10(-14) cm3 s-1 subunit-1) and low Arrhenius activation energy (Ea = 3.7 kcal/mol), similar to mammalian aquaporin-1 (AQP1). No permeation by glycerol, urea or sorbitol was detected. Expression of native and modified AqpZ in milligram quantities has permitted biophysical characterization of this remarkably stable aquaporin tetramer, which is being utilized for high-resolution structural studies.
</noinclude>
|-
|{{TableHeadingColor}} nowrap |'''Links''' || [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10518952 PubMed]
|}
</protect>
==Significance {{HelpIcon|Help:Literature}} ==
{{LitSignificance}}

== Useful Materials and Methods {{HelpIcon|Help:Literature}}==
{{LitMaterials}}

==Annotations {{HelpIcon|Help:Literature}}==
<protect>

{|class = 'sortable' {{Prettytable}} id='239708'
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!Gene product!!Qualifier!!GO ID!!GO term name!!Evidence Code!!with/from!!Aspect!!Notes!!Status
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</protect>

==EcoliWiki Links {{HelpIcon|Help:Literature}} ==
{{LitLinks}}

==References==
{{RefHelp}}
<references/>