TY  - JOUR
AU  - Kriegsmann, J.
AU  - Gregor, I.
AU  - von der Hocht, I.
AU  - Klare, J.
AU  - Engelhard, M.
AU  - Enderlein, J.
AU  - Fitter, J.
TI  - Translational Diffusion and Interaction of a Photoreceptor and Its Cognate Transducer Observed in Giant Unilamellar Vesicles by Using Dual-Focus FCS
JO  - ChemBioChem
VL  - 10
SN  - 1439-4227
CY  - Weinheim
PB  - Wiley-VCH
M1  - PreJuSER-5165
SP  - 1823 - 1829
PY  - 2009
N1  - The priority program SPP 7128 of Deutsche Forschungsgemeinschaft (F1 84113-1,2 to J.F and to J.E) and the DFG graduate school 1035 "Biointerface" are acknowledged for funding. We thank B. Kaupp and G. Baldt for continuous support in their institutes.
AB  - In order to monitor membrane-protein binding in lipid bilayers at physiological protein concentrations, we employed the recently developed dual-focus fluorescence correlation spectroscopy (2fFCS) technique. In a case study on a photoreceptor consisting of seven transmembrane helices and its cognate transducer (two transmembrane helices), the lateral diffusion for these integral membrane proteins was analyzed in giant unilamellar vesicles (GUVs). The two-dimensional diffusion coefficients of both separately diffusing proteins differ significantly, with D = 2.2 x 10(-8) cm2 s(-1) for the photoreceptor and with D = 4.1 x 10(-8) cm2 s(-1) for the transducer. In GUVs with both membrane proteins present together, we observed significantly smaller diffusion coefficients for labelled transducer molecules; this indicates the presence of larger diffusing units and therefore intermolecular protein binding. Based on the phenomenological dependence of diffusion coefficients on the molecule's cylindrical radius, we are able to estimate the degree of membrane protein binding on a quantitative level.
KW  - Archaeal Proteins: chemistry
KW  - Archaeal Proteins: metabolism
KW  - Carotenoids: chemistry
KW  - Carotenoids: metabolism
KW  - Diffusion
KW  - Lipid Bilayers: chemistry
KW  - Membrane Proteins: chemistry
KW  - Photoreceptors, Microbial: chemistry
KW  - Photoreceptors, Microbial: metabolism
KW  - Protein Binding
KW  - Spectrometry, Fluorescence
KW  - Unilamellar Liposomes: chemistry
KW  - Archaeal Proteins (NLM Chemicals)
KW  - HtrII protein, Natronobacterium pharaonis (NLM Chemicals)
KW  - Lipid Bilayers (NLM Chemicals)
KW  - Membrane Proteins (NLM Chemicals)
KW  - Photoreceptors, Microbial (NLM Chemicals)
KW  - Unilamellar Liposomes (NLM Chemicals)
KW  - phototaxis receptor sensory rhodopsin II, Natronobacterium pharaonis (NLM Chemicals)
KW  - Carotenoids (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:19551796
UR  - <Go to ISI:>//WOS:000268504500015
DO  - DOI:10.1002/cbic.200900251
UR  - https://juser.fz-juelich.de/record/5165
ER  -