schliessen

Filtern

 

Bibliotheken

Cell-free expression and in meso crystallisation of an integral membrane kinase for structure determination

Membrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when s... Full description

Journal Title: Cellular and Molecular Life Sciences 2014, Vol.71(24), pp.4895-4910
Main Author: Boland, Coilín
Other Authors: Li, Dianfan , Shah, Syed , Haberstock, Stefan , Dötsch, Volker , Bernhard, Frank , Caffrey, Martin
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 1420-682X ; E-ISSN: 1420-9071 ; DOI: 10.1007/s00018-014-1655-7
Link: http://dx.doi.org/10.1007/s00018-014-1655-7
Zum Text:
SendSend as email Add to Book BagAdd to Book Bag
Staff View
recordid: springer_jour10.1007/s00018-014-1655-7
title: Cell-free expression and in meso crystallisation of an integral membrane kinase for structure determination
format: Article
creator:
  • Boland, Coilín
  • Li, Dianfan
  • Shah, Syed
  • Haberstock, Stefan
  • Dötsch, Volker
  • Bernhard, Frank
  • Caffrey, Martin
subjects:
  • Cell-free
  • Crystallisation
  • In meso
  • Lipid cubic phase
  • Macromolecular crystallography
  • Membrane protein structure
ispartof: Cellular and Molecular Life Sciences, 2014, Vol.71(24), pp.4895-4910
description: Membrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.
language: eng
source:
identifier: ISSN: 1420-682X ; E-ISSN: 1420-9071 ; DOI: 10.1007/s00018-014-1655-7
fulltext: fulltext
issn:
  • 1420-9071
  • 14209071
  • 1420-682X
  • 1420682X
url: Link


@attributes
ID1090497956
RANK0.07
NO1
SEARCH_ENGINEprimo_central_multiple_fe
SEARCH_ENGINE_TYPEPrimo Central Search Engine
LOCALfalse
PrimoNMBib
record
control
sourcerecordid10.1007/s00018-014-1655-7
sourceidspringer_jour
recordidTN_springer_jour10.1007/s00018-014-1655-7
sourcesystemOther
pqid1626164692
galeid390294570
display
typearticle
titleCell-free expression and in meso crystallisation of an integral membrane kinase for structure determination
creatorBoland, Coilín ; Li, Dianfan ; Shah, Syed ; Haberstock, Stefan ; Dötsch, Volker ; Bernhard, Frank ; Caffrey, Martin
ispartofCellular and Molecular Life Sciences, 2014, Vol.71(24), pp.4895-4910
identifier
subjectCell-free ; Crystallisation ; In meso ; Lipid cubic phase ; Macromolecular crystallography ; Membrane protein structure
descriptionMembrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.
languageeng
source
version8
lds50peer_reviewed
links
openurl$$Topenurl_article
openurlfulltext$$Topenurlfull_article
backlink$$Uhttp://dx.doi.org/10.1007/s00018-014-1655-7$$EView_full_text_in_Springer_(Subscribers_only)
search
creatorcontrib
0Boland, Coilín, Tasadaque
1Li, Dianfan, Tasadaque
2Shah, Syed, Tasadaque
3Haberstock, Stefan, Tasadaque
4Dötsch, Volker, Tasadaque
5Bernhard, Frank, Tasadaque
6Caffrey, Martin, Tasadaque
titleCell-free expression and in meso crystallisation of an integral membrane kinase for structure determination
descriptionMembrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.
subject
0Cell-free
1Crystallisation
2In meso
3Lipid cubic phase
4Macromolecular crystallography
5Membrane protein structure
general
010.1007/s00018-014-1655-7
1English
2Springer Science & Business Media B.V.
3SpringerLink
sourceidspringer_jour
recordidspringer_jour10.1007/s00018-014-1655-7
issn
01420-9071
114209071
21420-682X
31420682X
rsrctypearticle
creationdate2014
addtitle
0Cellular and Molecular Life Sciences
1Cell. Mol. Life Sci.
searchscopespringer_journals_complete
scopespringer_journals_complete
lsr30VSR-Enriched:[pqid, pages, galeid]
sort
titleCell-free expression and in meso crystallisation of an integral membrane kinase for structure determination
authorBoland, Coilín ; Li, Dianfan ; Shah, Syed ; Haberstock, Stefan ; Dötsch, Volker ; Bernhard, Frank ; Caffrey, Martin
creationdate20141200
facets
frbrgroupid4906399313340890088
frbrtype5
languageeng
creationdate2014
topic
0Cell-Free
1Crystallisation
2In Meso
3Lipid Cubic Phase
4Macromolecular Crystallography
5Membrane Protein Structure
collectionSpringerLink
prefilterarticles
rsrctypearticles
creatorcontrib
0Boland, Coilín
1Li, Dianfan
2Shah, Syed
3Haberstock, Stefan
4Dötsch, Volker
5Bernhard, Frank
6Caffrey, Martin
jtitleCellular And Molecular Life Sciences
toplevelpeer_reviewed
delivery
delcategoryRemote Search Resource
fulltextfulltext
addata
aulast
0Boland
1Li
2Shah
3Haberstock
4Dötsch
5Bernhard
6Caffrey
aufirst
0Coilín
1Dianfan
2Syed
3Tasadaque
4Ali
5Stefan
6Volker
7Frank
8Martin
au
0Boland, Coilín
1Li, Dianfan
2Shah, Syed
3Haberstock, Stefan
4Dötsch, Volker
5Bernhard, Frank
6Caffrey, Martin
atitleCell-free expression and in meso crystallisation of an integral membrane kinase for structure determination
jtitleCellular and Molecular Life Sciences
stitleCell. Mol. Life Sci.
risdate201412
volume71
issue24
spage4895
epage4910
issn1420-682X
eissn1420-9071
genrearticle
ristypeJOUR
abstractMembrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.
copBasel
pubSpringer Basel
doi10.1007/s00018-014-1655-7
pages4895-4910
date2014-12