TY - JOUR
T1 - Spectroscopic and kinetic characterization of eosin-5-maleimide
AU - Schopfer, Lawrence M.
AU - Salhany, James M.
N1 - Funding Information:
1This work was supported by the Medical Service of the Veterans Administration. 2 To whom correspondence should be addressed at the Department of Internal Medicine, University of Nebraska Medical Center, 600 South 42nd Street, Omaha, NE 68198-5290. Fax: (402) 559-9004. 3Abbreviations used: EM, eosin–5-maleimide; EMA, eosin–5-maleic acid; Mes, 2-(N-morpholino)ethane sulfonic acid; DMF, dimethylformamide; DMSO, dimethylsulfoxide; C12E8, polyoxyethylene-8-lauryl ether; FL, fluorescein.
PY - 1998/3/15
Y1 - 1998/3/15
N2 - Eosin-5-maleimide (EM) is an increasingly important and widely used probe in the study of membrane protein structure and function. Yet little is known about its spectral properties in hydrophobic and hydrophilic environments. Furthermore, EM is hydrolyzed faster than the traditional N- ethylmaleimide. To offer a more solid foundation for the use of EM in studies of membrane protein structure and function, we have undertaken a detailed study of the absorbance and fluorescence spectra of EM, eosin-5-maleic acid, and the L-cysteine and β-mercaptoethanol adducts of EM in aqueous and hydrophobic environments; we have studied the kinetics of hydrolysis of EM in various environments, and we have investigated the reaction kinetics of EM with L-cysteine.
AB - Eosin-5-maleimide (EM) is an increasingly important and widely used probe in the study of membrane protein structure and function. Yet little is known about its spectral properties in hydrophobic and hydrophilic environments. Furthermore, EM is hydrolyzed faster than the traditional N- ethylmaleimide. To offer a more solid foundation for the use of EM in studies of membrane protein structure and function, we have undertaken a detailed study of the absorbance and fluorescence spectra of EM, eosin-5-maleic acid, and the L-cysteine and β-mercaptoethanol adducts of EM in aqueous and hydrophobic environments; we have studied the kinetics of hydrolysis of EM in various environments, and we have investigated the reaction kinetics of EM with L-cysteine.
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U2 - 10.1006/abio.1998.2575
DO - 10.1006/abio.1998.2575
M3 - Article
C2 - 9514795
AN - SCOPUS:0032521048
SN - 0003-2697
VL - 257
SP - 139
EP - 148
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -