TY  - JOUR
AU  - Skegro, D.
AU  - Pulvermuller, A.
AU  - Krafft, B.
AU  - Granzin, J.
AU  - Hofmann, K. P.
AU  - Büldt, G.
AU  - Schlesinger, R.
TI  - N-terminal and C-terminal Domains of Arrestin Both Contribute in Binding to Rhodopsin (dagger)
JO  - Photochemistry and photobiology
VL  - 83
SN  - 0031-8655
CY  - Malden, Mass.
PB  - Wiley-Blackwell
M1  - PreJuSER-57375
SP  - 385 - 393
PY  - 2007
N1  - Record converted from VDB: 12.11.2012
AB  - Visual arrestin terminates the signal amplification cascade in photoreceptor cells by blocking the interaction of light activated phosphorylated rhodopsin with the G-protein transducin. Although crystal structures of arrestin and rhodopsin are available, it is still unknown how the complex of the two proteins is formed. To investigate the interaction sites of arrestin with rhodopsin various surface regions of recombinant arrestin were sterically blocked by different numbers of fluorophores (Alexa 633). The binding was recorded by time-resolved light scattering. To accomplish site-specific shielding of protein regions, in a first step all three wild-type cysteines were replaced by alanines. Nevertheless, regarding the magnitude and specificity of rhodopsin binding, the protein is still fully active. In a second step, new cysteines were introduced at selected sites to allow covalent binding of fluorophores. Upon attachment of Alexa 633 to the recombinant cysteines we observed that these bulky labels residing in the concave area of either the N- or the C-terminal domain do not perturb the activity of arrestin. By simultaneously modifying both domains with one Alexa 633 the binding capacity was reduced. The presence of two Alexa 633 molecules in each domain prevented binding of rhodopsin to arrestin. This observation indicates that both concave sites participate in binding.
KW  - Animals
KW  - Arrestin: chemistry
KW  - Arrestin: genetics
KW  - Arrestin: metabolism
KW  - Base Sequence
KW  - Binding Sites
KW  - Cattle
KW  - Cysteine: chemistry
KW  - DNA Primers: genetics
KW  - Fluorescent Dyes
KW  - Models, Molecular
KW  - Mutagenesis, Site-Directed
KW  - Photochemistry
KW  - Protein Binding
KW  - Protein Structure, Tertiary
KW  - Recombinant Proteins: chemistry
KW  - Recombinant Proteins: genetics
KW  - Recombinant Proteins: metabolism
KW  - Rhodopsin: metabolism
KW  - Signal Transduction
KW  - Arrestin (NLM Chemicals)
KW  - DNA Primers (NLM Chemicals)
KW  - Fluorescent Dyes (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
KW  - Cysteine (NLM Chemicals)
KW  - Rhodopsin (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:17132044
UR  - <Go to ISI:>//WOS:000245658000025
DO  - DOI:10.1562/2006-08-25-RA-1014
UR  - https://juser.fz-juelich.de/record/57375
ER  -