-
Something wrong with this record ?
Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of AHP2, a signal transmitter protein from Arabidopsis thaliana
O. Degtjarik, R. Dopitova, S. Puehringer, E. Nejedla, M. Kuty, MS. Weiss, J. Hejatko, L. Janda, I. Kuta Smatanova,
Language English Country England, Great Britain
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
Free Medical Journals
from 2005 to 2013
PubMed Central
from 2005 to 2013
Europe PubMed Central
from 2005 to 2013
- MeSH
- Arabidopsis metabolism MeSH
- X-Ray Diffraction MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Phosphotransferases chemistry isolation & purification MeSH
- Crystallization MeSH
- Arabidopsis Proteins chemistry isolation & purification MeSH
- Signal Transduction * MeSH
- Transition Temperature MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Histidine-containing phosphotransfer proteins from Arabidopsis thaliana (AHP1-5) act as intermediates between sensor histidine kinases and response regulators in a signalling system called multi-step phosphorelay (MSP). AHP proteins mediate and potentially integrate various MSP-based signalling pathways (e.g. cytokinin or osmosensing). However, structural information about AHP proteins and their importance in MSP signalling is still lacking. To obtain a deeper insight into the structural basis of AHP-mediated signal transduction, the three-dimensional structure of AHP2 was determined. The AHP2 coding sequence was cloned into pRSET B expression vector, enabling production of AHP2 fused to an N-terminal His tag. AHP2 was expressed in soluble form in Escherichia coli strain BL21 (DE3) pLysS and then purified to homogeneity using metal chelate affinity chromatography and anion-exchange chromatography under reducing conditions. Successful crystallization in a buffer which was optimized for thermal stability yielded crystals that diffracted to 2.5 Å resolution.
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc13031640
- 003
- CZ-PrNML
- 005
- 20131002124447.0
- 007
- ta
- 008
- 131002s2013 enk f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1107/S174430911205186X $2 doi
- 035 __
- $a (PubMed)23385758
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a enk
- 100 1_
- $a Degtjarik, Oksana $u South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses and School of Complex Systems, University of South Bohemia, Zamek 136, 37333 Nove Hrady, Czech Republic.
- 245 10
- $a Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of AHP2, a signal transmitter protein from Arabidopsis thaliana / $c O. Degtjarik, R. Dopitova, S. Puehringer, E. Nejedla, M. Kuty, MS. Weiss, J. Hejatko, L. Janda, I. Kuta Smatanova,
- 520 9_
- $a Histidine-containing phosphotransfer proteins from Arabidopsis thaliana (AHP1-5) act as intermediates between sensor histidine kinases and response regulators in a signalling system called multi-step phosphorelay (MSP). AHP proteins mediate and potentially integrate various MSP-based signalling pathways (e.g. cytokinin or osmosensing). However, structural information about AHP proteins and their importance in MSP signalling is still lacking. To obtain a deeper insight into the structural basis of AHP-mediated signal transduction, the three-dimensional structure of AHP2 was determined. The AHP2 coding sequence was cloned into pRSET B expression vector, enabling production of AHP2 fused to an N-terminal His tag. AHP2 was expressed in soluble form in Escherichia coli strain BL21 (DE3) pLysS and then purified to homogeneity using metal chelate affinity chromatography and anion-exchange chromatography under reducing conditions. Successful crystallization in a buffer which was optimized for thermal stability yielded crystals that diffracted to 2.5 Å resolution.
- 650 _2
- $a Arabidopsis $x metabolismus $7 D017360
- 650 _2
- $a proteiny huseníčku $x chemie $x izolace a purifikace $7 D029681
- 650 _2
- $a krystalizace $7 D003460
- 650 _2
- $a elektroforéza v polyakrylamidovém gelu $7 D004591
- 650 _2
- $a fosfotransferasy $x chemie $x izolace a purifikace $7 D010770
- 650 12
- $a signální transdukce $7 D015398
- 650 _2
- $a tranzitní teplota $7 D044366
- 650 _2
- $a difrakce rentgenového záření $7 D014961
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Dopitova, Radka $u -
- 700 1_
- $a Puehringer, Sandra $u -
- 700 1_
- $a Nejedla, Eliska $u -
- 700 1_
- $a Kuty, Michal $u -
- 700 1_
- $a Weiss, Manfred S $u -
- 700 1_
- $a Hejatko, Jan $u -
- 700 1_
- $a Janda, Lubomir $u -
- 700 1_
- $a Kuta Smatanova, Ivana $u -
- 773 0_
- $w MED00179506 $t Acta crystallographica. Section F, Structural biology and crystallization communications $x 1744-3091 $g Roč. 69, č. Pt 2 (2013), s. 158-61
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/23385758 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20131002 $b ABA008
- 991 __
- $a 20131002125004 $b ABA008
- 999 __
- $a ok $b bmc $g 995727 $s 830085
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2013 $b 69 $c Pt 2 $d 158-61 $i 1744-3091 $m Acta crystallographica. Section F $n Acta Crystallogr Sect F Struct Biol Cryst Commun $x MED00179506
- LZP __
- $a Pubmed-20131002
