Dynamics surrounding Cys-34 in native, chemically denatured, and silica-adsorbed bovine serum albumin.
We report the steady-state and time-resolved fluorescence of 6-acryloyl(dimethylamino)naphthalene (acrylodan) covalently attached to Cys-34 in bovine serum albumin (BSA). For this conceptually simple system, complicated fluorescence intensity and anisotropy decay kinetics are observed. The steady-state and time-resolved results demonstrate the presence of an excited-state reaction for the BSA-acrylodan system. Additional analysis shows that dipolar relaxation of the environment surrounding acrylodan within BSA is responsible for most of the observed time-dependent evolution of the emission spectrum. The effects of temperature, chemical denaturation, and protein adsorption to a bare silica substrate are also investigated. These results demonstrate the complexity of the changes within a protein/biorecognition element that affect the signal from a single fluorescent reporter group.
Wang
R
R
Department of Chemistry, Natural Sciences and Mathematics Complex, State University of New York at Buffalo 14260-3000.
Sun
S
S
Bekos
E J
EJ
Bright
F V
FV
D015081:2-Naphthylamine / Q000031:analogs & derivatives / Q000737:chemistry; D000595:Amino Acid Sequence; D003545:Cysteine / Q000032:analysis*; D005454:Fluorescence Polarization; D007700:Kinetics; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012822:Silicon Dioxide; D013997:Time Factors
D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; C038518:acrylodan; D015081:2-Naphthylamine; D003545:Cysteine