Fluorescein-labeled glutathione to study protein S-glutathionylation.
Numerous studies of S-glutathionylation of cysteine thiols indicate that this protein modification plays a key role in redox regulation of proteins. To facilitate the study of protein S-glutathionylation, we developed a synthesis and purification to produce milligram quantities of fluorescein-labeled glutathione. The amino terminus of the glutathione tripeptide reacted with fluorescein isothiocyanate readily in ammonium bicarbonate. Purification by solid phase extraction on C8 and C18 columns separated excess reactants from desired products. Both oxidized and reduced fluorescein-labeled glutathione reacted with a variety of thiol-containing proteins to yield fluorescent proteins.
Landino
Lisa M
LM
Department of Chemistry, College of William and Mary, Williamsburg, VA 23187, USA. lmland@wm.edu
Brown
Carolyn M
CM
Edson
Carolyn A
CA
Gilbert
Laura J
LJ
Grega-Larson
Nathan
N
Wirth
Anna Jean
AJ
Lane
Kelly C
KC
D019793:Fluorescein / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D011506:Proteins / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D005456:Fluorescent Dyes; D011506:Proteins; D013438:Sulfhydryl Compounds; D005978:Glutathione; D019793:Fluorescein