A major fraction of endoplasmic reticulum-located glutathione is present as mixed disulfides with protein.
The tripeptide glutathione is the most abundant thiol/disulfide component of the eukaryotic cell and is known to be present in the endoplasmic reticulum lumen. Accordingly, the thiol/disulfide redox status of the endoplasmic reticulum lumen is defined by the status of glutathione, and it has been assumed that reduced and oxidized glutathione form the principal redox buffer. We have determined the distribution of glutathione between different chemical states in rat liver microsomes by labeling with the thiol-specific label monobromobimane and subsequent separation by reversed phase high performance liquid chromatography. More than half of the microsomal glutathione was found to be present in mixed disulfides with protein, the remainder being distributed between the reduced and oxidized forms of glutathione in the ratio of 3:1. The high proportion of the total population of glutathione that was found to be in mixed disulfides with protein has significant implications for the redox state and buffering capacity of the endoplasmic reticulum and, hence, for the formation of disulfide bonds in vivo.
Bass
Rosemary
R
Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, United Kingdom.
Ruddock
Lloyd W
LW
Klappa
Peter
P
Freedman
Robert B
RB
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000737:chemistry; D008099:Liver / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D051381:Rats; D013997:Time Factors
D004220:Disulfides; D010455:Peptides; D005978:Glutathione; D010100:Oxygen; D019803:Glutathione Disulfide