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.
Department of Chemistry, College of William and Mary, Williamsburg, VA 23187, USA. lmland@wm.edu
Lisa M
LM
Landino
Carolyn M
CM
Brown
Carolyn A
CA
Edson
Laura J
LJ
Gilbert
Nathan
N
Grega-Larson
Anna Jean
AJ
Wirth
Kelly C
KC
Lane
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