TY  - JOUR
AU  - Mennes, N.
AU  - Klare, J. P.
AU  - Chizhov, I.
AU  - Seidel, R.
AU  - Schlesinger, R.
AU  - Engelhard, M.
TI  - Expression of the halobacterial transducer protein HtrII from Natronomonas pharaonis in Escherichia coli
JO  - FEBS letters
VL  - 581
SN  - 0014-5793
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - PreJuSER-57377
SP  - 1487 - 1494
PY  - 2007
N1  - Record converted from VDB: 12.11.2012
AB  - Archaeal phototaxis is mediated by sensory rhodopsins which form complexes with their cognate transducers. Whereas the receptors sensory rhodopsin I and sensory rhodopsin II (SRII) have been expressed in Escherichia coli (E. coli) only shortened fragments of HtrII from Natronomonas pharaonis (NpHtrII) are available. Here we describe the heterologous expression of full length NpHtrII which was achieved in yields of up to 0.9 mg per litre cell culture. Gel filtration analysis reveals the tendency of the transducer to form dimers and higher-order oligomers which was also observed when complexed to NpSRII. A circular dichroism (CD) spectrum of NpHtrII is comparable to those obtained for the E. coli chemoreceptors indicating a similar folding with predominantly alpha-helical structure. NpHtrII dissociates from the NpSRII/HtrII complex with an apparent K(D) of about 0.6 microM. Photocycle kinetics of the complex is comparable to that obtained for NpSRII in complex with a truncated transducer with slight differences in the M-decay. The data indicate that the heterologously expressed NpHtrII adopt a native like structure, providing the means for elucidating transmembrane signal transduction and activation of microbial signalling cascades.
KW  - Archaeal Proteins: biosynthesis
KW  - Archaeal Proteins: chemistry
KW  - Archaeal Proteins: isolation & purification
KW  - Chromatography, Gel
KW  - Circular Dichroism
KW  - Cloning, Molecular
KW  - Dimerization
KW  - Escherichia coli: genetics
KW  - Lasers
KW  - Photolysis
KW  - Protein Structure, Secondary
KW  - Recombinant Proteins: biosynthesis
KW  - Recombinant Proteins: chemistry
KW  - Recombinant Proteins: isolation & purification
KW  - Archaeal Proteins (NLM Chemicals)
KW  - HtrII protein, Natronobacterium pharaonis (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:17368449
UR  - <Go to ISI:>//WOS:000245913300036
DO  - DOI:10.1016/j.febslet.2007.03.005
UR  - https://juser.fz-juelich.de/record/57377
ER  -