Characterization of the interactions of fluorescent probes with proteins: coumarin 153 and 1,8-ANS in complex with holo- and apomyoglobin.
In order to provide a thorough characterization of a system with which to study the dielectric response of a protein, a well-defined system complex of a fluorescent probe and protein is required. We have argued that such a system is provided by coumarin 153 and apomyoglobin (Photochem. Photobiol. 79, 440-446 [2004]). In order to demonstrate further that coumarin 153 exhibits negligible nonspecific binding to the surface of apomyoglobin, we study its interactions with both the apo and holo proteins. We further make a similar comparison with 8-anilino-l-naphthalenesulfonic acid, for which an NMR structure with apomyoglobin has been obtained. Our results confirm the appropriateness of the system of coumarin 153 and apomyoglobin for the investigation of solvation by the protein matrix.
Mukherjee
Prasun
P
Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
Halder
Mintu
M
Hargrove
Mark S
MS
Petrich
Jacob W
JW
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry*; D000818:Animals; D001059:Apoproteins / Q000737:chemistry*; D003374:Coumarins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006736:Horses; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D009211:Myoglobin / Q000737:chemistry*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
D000817:Anilino Naphthalenesulfonates; D001059:Apoproteins; D003374:Coumarins; D005456:Fluorescent Dyes; D009211:Myoglobin; C010857:apomyoglobin; C488877:coumarin 153; C027132:1-anilino-8-naphthalenesulfonate