TY - JOUR
AU - Kriegsmann, J.
AU - Brehs, M.
AU - Klare, J.P.
AU - Engelhard, M.
AU - Fitter, J.
TI - Sensory rhodopsin II/transducer complex formation in detergent and in lipid bilayers studied with FRET
JO - Biochimica et biophysica acta / Biomembranes
VL - 1788
SN - 0005-2736
CY - Amsterdam
PB - Elsevier
M1 - PreJuSER-5168
SP - 522 - 531
PY - 2009
N1 - J. F. thanks G. Buldt (Forschungszentrum Julich) for continuous and sustainable support in his institute. This work was supported by the Deutsche Forschungsgerneinschaft (M.E.) and by a priority program SPP 1128 of Deutsche Forschungsgerneinschaft (FI 841/3-1,2 to J.F.). We are indebted to R. Schlesinger for providing us with bacteriorhodopsin and to R. Batra-Safferling for the discussions and comments on the manuscript.
AB - The photophobic receptor from Natronomonas pharaonis (NpSRII) forms a photo-signalling complex with its cognate transducer (NpHtrII). In order to elucidate the complex formation in more detail, we have studied the intermolecular binding of both constituents (NpSRII and NpHtrII157; truncated at residue 157) in detergent buffers, and in lipid bilayers using FRET. The data for hetero-dimer formation of NpSRII/NpHtrII in detergent agrees well with KD values (approximately 200 nM) described in the literature. In lipid bilayers, the binding affinity between proteins in the NpSRII/NpHtrII complex is at least one order of magnitude stronger. In detergent the strength of binding is similar for both homo-dimers (NpSRII/NpSRII and NpHtrII/NpHtrII) but significantly weaker (KD approximately 16 microM) when compared to the hetero-dimer. The intermolecular binding is again considerably stronger in lipid bilayers; however, it is not as strong as that observed for the hetero-dimer. At a molar transducer/lipid ratio of 1:2000, which is still well above physiological concentrations, only 40% homo-dimers are formed. Apparently, in cell membranes the formation of the assumed functionally active oligomeric 2:2 complex depends on the full-length transducer including the helical cytoplasmic part, which is thought to tighten the transducer-dimer association.
KW - Archaeal Proteins: chemistry
KW - Archaeal Proteins: genetics
KW - Archaeal Proteins: metabolism
KW - Biophysical Phenomena
KW - Detergents
KW - Halobacteriaceae: chemistry
KW - Halobacteriaceae: genetics
KW - Halobacteriaceae: metabolism
KW - Lipid Bilayers: chemistry
KW - Models, Molecular
KW - Protein Binding
KW - Protein Multimerization
KW - Protein Structure, Quaternary
KW - Sensory Rhodopsins: chemistry
KW - Sensory Rhodopsins: genetics
KW - Sensory Rhodopsins: metabolism
KW - Signal Transduction
KW - Spectrophotometry
KW - Archaeal Proteins (NLM Chemicals)
KW - Detergents (NLM Chemicals)
KW - HtrII protein, Natronobacterium pharaonis (NLM Chemicals)
KW - Lipid Bilayers (NLM Chemicals)
KW - Sensory Rhodopsins (NLM Chemicals)
KW - J (WoSType)
LB - PUB:(DE-HGF)16
C6 - pmid:19094962
UR - <Go to ISI:>//WOS:000263794700025
DO - DOI:10.1016/j.bbamem.2008.11.011
UR - https://juser.fz-juelich.de/record/5168
ER -