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@ARTICLE{Mennes:57377,
author = {Mennes, N. and Klare, J. P. and Chizhov, I. and Seidel, R.
and Schlesinger, R. and Engelhard, M.},
title = {{E}xpression of the halobacterial transducer protein
{H}tr{II} from {N}atronomonas pharaonis in {E}scherichia
coli},
journal = {FEBS letters},
volume = {581},
issn = {0014-5793},
address = {Amsterdam [u.a.]},
publisher = {Elsevier},
reportid = {PreJuSER-57377},
pages = {1487 - 1494},
year = {2007},
note = {Record converted from VDB: 12.11.2012},
abstract = {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.},
keywords = {Archaeal Proteins: biosynthesis / Archaeal Proteins:
chemistry / Archaeal Proteins: isolation $\&$ purification /
Chromatography, Gel / Circular Dichroism / Cloning,
Molecular / Dimerization / Escherichia coli: genetics /
Lasers / Photolysis / Protein Structure, Secondary /
Recombinant Proteins: biosynthesis / Recombinant Proteins:
chemistry / Recombinant Proteins: isolation $\&$
purification / Archaeal Proteins (NLM Chemicals) / HtrII
protein, Natronobacterium pharaonis (NLM Chemicals) /
Recombinant Proteins (NLM Chemicals) / J (WoSType)},
cin = {INB-2},
ddc = {570},
cid = {I:(DE-Juel1)VDB805},
pnm = {Funktion und Dysfunktion des Nervensystems},
pid = {G:(DE-Juel1)FUEK409},
shelfmark = {Biochemistry $\&$ Molecular Biology / Biophysics / Cell
Biology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:17368449},
UT = {WOS:000245913300036},
doi = {10.1016/j.febslet.2007.03.005},
url = {https://juser.fz-juelich.de/record/57377},
}