The Utility of N,N-Biotinyl Glutathione Disulfide in the Study of Protein S-Glutathiolation

Abstract

Glutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or regulate protein function. We developed novel strategies for the study of protein S-glutathiolation that involved the simple synthesis of N,N-biotinyl glutathione disulfide (biotin-GSSG). Biotin-GSSG of cells a of a in the glutathione the disulfide protein S-glutathiolation, which We that a in the of protein S-glutathiolation or novel in the study of disease or GSSG Glutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or regulate protein function. We developed novel strategies for the study of protein S-glutathiolation that involved the simple synthesis of N,N-biotinyl glutathione disulfide (biotin-GSSG). Biotin-GSSG of cells a of a in the glutathione the disulfide protein S-glutathiolation, which We that a in the of protein S-glutathiolation or novel in the study of disease or GSSG in the of in the of in of cells the of protein a of which or of as S-glutathiolation, a protein in a the well of of protein S-glutathiolation in protein a of the of the of the the in which the a which the of the of protein S-glutathiolation in of for of many a in the glutathione N,N-biotinyl glutathione glutathione N,N-biotinyl glutathione a in glutathione in the of glutathione in of glutathione in in glutathione the in in cells in of in the of the of an the GSSG accumulates in can disulfide S-glutathiolation, which can as the that protein S-glutathiolation in disease in the of for novel for the study of protein S-glutathiolation in which GSSG N,N-biotinyl glutathione disulfide We as for the the We as a for that of a of a or under an in in GSSG protein of that that a which that a of that in the the of the of the of glutathione disulfide under the GSSG in the for a of the an a that in a an a of N,N-biotinyl glutathione the of which as the of of the N,N-biotinyl glutathione disulfide for an of for of the disulfide for protein in of of in a Biotin-GSSG of a of of the for for the of S-glutathiolation, the in of or during a of of as a GSSG as an or GSSG for or the in for protein S-glutathiolation We in as of a protein of a as of in for GSSG disulfide in cells for GSSG or cells for the in for for the in in the in in of for a of of cells for the in in of as for a of or an of as in of the of or in a in of that S-glutathiolation GSSG or a the in which of GSSG or as protein the of the of in the the of the GSSG or a a a of of GSSG in the of the in the as a the in a of disulfide the a protein for of in of of for a of protein the the of in for a of a in a the an of or a of a a as of in the of the of or of as the of of the for that a or that the of the of GSSG or as for cells the the cells the the in in cells or of a cells a cells a of an the of a the for of of an in of the of S-glutathiolation in a of protein in of a an protein S-glutathiolation in of under or GSSG or protein S-glutathiolation in the a in the of of protein in the GSSG S-glutathiolation the that the involved disulfide which the of the as a which protein promotes S-glutathiolation an or in the study of in cells directly protein S-glutathiolation or the of GSSG can protein a that S-glutathiolation of in cells that GSSG or in the the protein S-glutathiolation in the GSSG that in reversible of in for S-glutathiolation a in the or of a of in a that in the of a in for S-glutathiolation for the of in the of S-glutathiolation in a of S-glutathiolation in the of of the of in that of a S-glutathiolation a of of the of during oxidant the of in a for a that an that in of the GSSG the of the or in the the in a of the increased the of as the in a the in S-glutathiolation as a a of protein in of cells the of protein S-glutathiolation a GSSG in that GSSG the of that that S-glutathiolation of the protein as or a of GSSG S-glutathiolation, that the protein the for of or cells for the of a of or the the of a the of a protein the of or the of the that the protein of of We of the many of the the in the metabolic as in the study of in cells of of of protein of of can in of protein a protein protein of of of in a protein the protein protein in a the of the the as protein of the of that can in protein protein protein in a of of in in that cells protein S-glutathiolation as the that in GSSG the an protein S-glutathiolation the the which the in the the of cells of the in the of in protein S-glutathiolation in as GSSG which the a the cells in a which in the the the disulfide (GSSG) of glutathione of the as the of the glutathione the of GSSG an of in many the a of an of in the disease can during the of cells or of the as the of the glutathione GSSG accumulates in protein in the of for of for the study of protein S-glutathiolation in in of that in of of the of that during of in the of a which of of of of that as the of can study of S-glutathiolation, the protein an of or in the study of protein of during of of during of during of protein of in protein in during which the for protein a of in the of in in or that a an of or metabolic or the of in of the which the oxidant a the oxidant many protein for the study of protein S-glutathiolation of the of many that the of the of glutathione in a of of can in the of as of Biotin-GSSG in the of of of the of of cells of an in as promotes protein S-glutathiolation, the the that of as well as of the that of protein S-glutathiolation, the of directly in of the that S-glutathiolation of in that for that S-glutathiolation in or that under of GSSG of S-glutathiolation an that GSSG the of a of the the of the the disulfide that protein the an the of the of which the the disulfide the that the of a of Biotin-GSSG that glutathione in for the of glutathione or the of of the of the of glutathione in of of can in that can for of in cells or as well as for for the of as that involved a protein in which the which in in S-glutathiolation of a of the of the of a of glutathione in that of the of protein S-glutathiolation the the the the of S-glutathiolation the of the of or for in the of that that the of that for S-glutathiolation the of in the a as the can as the of the of cells the the in which protein S-glutathiolation GSSG protein S-glutathiolation in cells the the of as a that the that which protein S-glutathiolation GSSG the of the the of GSSG as the promotes the cells in which the in the in cells GSSG of the in the or as of the of in of the a of of of the of a of that a for study of protein S-glutathiolation the of the study of of an as the disulfide that the in protein protein S-glutathiolation in or an the of for an S-glutathiolation as of S-glutathiolation of a of under in the of can in an or can cells or that directly in the disease or the in of the S-glutathiolation in of during during of during protein of of the metabolic or of in in that of that of S-glutathiolation that in of the the of the the of the of in the of that novel developed in the study of as GSSG a of many in the of in the of in of cells the of protein a of which or of as S-glutathiolation, a protein in a the well of of protein S-glutathiolation in protein a of the of the of the the in which the a which the of the of protein S-glutathiolation in of for of many a in the glutathione N,N-biotinyl glutathione glutathione N,N-biotinyl glutathione a in glutathione in the of glutathione in of glutathione in in glutathione the in in cells in of in the of the of an the GSSG accumulates in can disulfide S-glutathiolation, which can as the that protein S-glutathiolation in disease in the of for novel for the study of protein S-glutathiolation in which GSSG N,N-biotinyl glutathione disulfide We as for the the We as a for that of a of a or under an in in GSSG protein of that that a which that a of that in the the of the of the of glutathione disulfide under the GSSG in the for a of the an a that in a an a of N,N-biotinyl glutathione the of which as the of of the N,N-biotinyl glutathione disulfide for an of for of the disulfide for protein in of of in a Biotin-GSSG of a of of the for for the of S-glutathiolation, the in of or during a of of as a GSSG as an or GSSG for or the in for protein S-glutathiolation We in as of a protein of a as of in for GSSG disulfide in cells for GSSG or cells for the in for for the in in the in in of for a of of cells for the in in of as for a of or an of as in of the of or in a in of that S-glutathiolation GSSG or a the in which of GSSG or as protein the of the of in the the of the GSSG or a a a of of GSSG in the of the in the as a the in a of disulfide the a protein for of in of of for a of protein the the of in for a of a in a the an of or a of a a as of in the of the of or of as the of of the for that a or that the of the of GSSG or as for cells the the cells the the in in cells or of a cells a cells a of an the of a the for of in the the of the of the of glutathione disulfide under the GSSG in the for a of the an a that in a an a of N,N-biotinyl glutathione the of which as the of of the N,N-biotinyl glutathione disulfide for an of for of the disulfide for protein the of glutathione disulfide under the GSSG in the for a of the an a that in a an a of N,N-biotinyl glutathione the of which as the of of the N,N-biotinyl glutathione disulfide for an of for of the disulfide for protein in of of in a Biotin-GSSG of a of of the for for the of S-glutathiolation, the in of or during a of of as a GSSG as an or GSSG for or the in for protein S-glutathiolation We in as of a protein in of of in a Biotin-GSSG of a of of the for for the of S-glutathiolation, the in of or during a of of as a GSSG as an or GSSG for or the in for protein S-glutathiolation We in as of a protein of a as of in for of a as of in for GSSG disulfide in cells for GSSG or cells for the in for for the in in the in in of Glutathione disulfide in cells for GSSG or cells for the in for for the in in the in in of for a of of cells for the in in of as for a of or an of as in of for a of of cells for the in in of as for a of or an of as in of the of or in a in of that S-glutathiolation GSSG or a the in which the of or in a in of that S-glutathiolation GSSG or a the in which of GSSG or as protein the of the of in the the of the GSSG or a a a of of GSSG in the of the in the as a the in a of disulfide the a protein for GSSG or as protein the of the of in the the of the GSSG or a a a of of GSSG in the of the in the as a the in a of disulfide the a protein for of in of of for a of protein the the of in for a of a in a the an of or a of a a as of in the of the of or of as the of of the for that a or that the of the in of of for a of protein the the of in for a of a in a the an of or a of a a as of in the of the of or of as the of of the for that a or that the of the of GSSG or as for cells the the cells the the in in cells or of a cells a cells a of an the of a the for of GSSG or as for cells the the cells the the in in cells or of a cells a cells a of an the of a the for of of an in of the of S-glutathiolation in a of protein in of a an protein S-glutathiolation in of under or GSSG or protein S-glutathiolation in the a in the of of protein in the GSSG S-glutathiolation the that the involved disulfide which the of the as a which protein promotes S-glutathiolation an or in the study of in cells directly protein S-glutathiolation or the of GSSG can protein of in for S-glutathiolation a in the or of a of in a that in the of a in for S-glutathiolation for the of in the of S-glutathiolation in a of S-glutathiolation in the of of the of in that of a S-glutathiolation a of of the of during oxidant the of in a for a that an that in of the GSSG the of the or in the the in a of the increased the of as the in a the in S-glutathiolation as a a of protein in of cells the of protein S-glutathiolation a GSSG in that GSSG the of that that S-glutathiolation of the protein as or a of GSSG S-glutathiolation, that the protein the for of or cells for the of a of or the the of a the of a protein the of or the of the that the protein of of We of the many of the the in the metabolic as in the study of in cells of of of protein of of can in of protein a protein protein of of of in a protein the protein protein in a the of the the as protein of the of that can in protein protein protein in a of of in in that cells protein S-glutathiolation as the that in GSSG the an protein S-glutathiolation the the which the in the the of cells of the in the of in protein S-glutathiolation in as GSSG which the a the of an in of the of S-glutathiolation in a of protein in of a an protein S-glutathiolation in of under or GSSG or protein S-glutathiolation in the a in the of of protein in the GSSG S-glutathiolation the that the involved disulfide which the of the as a which protein promotes S-glutathiolation an or in the study of in cells directly protein S-glutathiolation or the of GSSG can protein of the of S-glutathiolation in a of protein in of a an protein S-glutathiolation in of under or GSSG or protein S-glutathiolation in the a in the of of protein in the GSSG S-glutathiolation the that the involved disulfide which the of the as a which protein promotes S-glutathiolation an or in the study of in cells directly protein S-glutathiolation or the of GSSG can protein of in for S-glutathiolation a in the or of a of in a that in the of a in for S-glutathiolation for the of in the of S-glutathiolation in a of for S-glutathiolation a in the or of a of in a that in the of a in for S-glutathiolation for the of in the of S-glutathiolation in a of of in that of a S-glutathiolation a of of the of during oxidant the of in a for a that an that in of the GSSG the of the or in the the in a of the increased the of as the the of protein S-glutathiolation a GSSG in that GSSG the of that that S-glutathiolation of the protein as or a of GSSG S-glutathiolation, that the protein the for of or cells for the that of a S-glutathiolation a of of the of during oxidant the of in a for a that an that in of the GSSG the of the or in the the in a of the increased the of as the the of protein S-glutathiolation a GSSG in that GSSG the of that that S-glutathiolation of the protein as of a of or the the of a the of a protein the protein of of We of the many of the the in the metabolic as in the study of in cells of of of protein of of can in of protein a protein protein of of of in a protein the protein protein in a the of the the as protein of the of that can in protein protein protein in a a of or the the of a the of a protein the protein of of We of the many of the the in the metabolic as in the study of in cells of of of protein of of can in of protein a protein protein of of of in a protein the protein protein in a the of the the as protein of the of that can in of of in in that cells protein S-glutathiolation as the that in GSSG the an protein S-glutathiolation the the which the in the the of cells of the in the of in in that cells protein S-glutathiolation as the that in GSSG the an protein S-glutathiolation the the which the in the the of cells of the in the cells in a which in the the the disulfide (GSSG) of glutathione of the as the of the glutathione the of GSSG an of in many the a of an of in the disease can during the of cells or of the as the of the glutathione GSSG accumulates in protein in the of for of for the study of protein S-glutathiolation in in of that in of of the of that during of in the of a which of of of of that as the of can study of S-glutathiolation, the protein an of or in the study of protein of during of of during of during of protein of in protein in during which the for protein a of in the of in in or that a an of or metabolic or the of in of the which the oxidant a the oxidant many protein for the study of protein S-glutathiolation of the of many that the of the of glutathione in a of of can in the of as of Biotin-GSSG in the of of of the of of cells of an in as promotes protein S-glutathiolation, the the that of as well as of the that of protein S-glutathiolation, the of directly in of the that S-glutathiolation of in that for that S-glutathiolation in or that under of GSSG of S-glutathiolation an that GSSG the of a of the the of the the disulfide that protein the an the of the of which the the disulfide the that the of a of Biotin-GSSG that glutathione in for the of glutathione or the of of the of the of glutathione in of of can in that can for of in cells or as well as for for the of as that involved a protein in which the which in in S-glutathiolation of a of the of the of a of glutathione in that of the of protein S-glutathiolation the the the the of S-glutathiolation the of the of or for in the of that that the of that for S-glutathiolation the of in the a as the can as the of the of cells the the in which protein S-glutathiolation GSSG protein S-glutathiolation in cells the the of as a that the that which protein S-glutathiolation GSSG the of the the of GSSG as the promotes the cells in which the in the in cells GSSG of the in the or as of the of in of the a of of of the of a of that a for study of protein S-glutathiolation the of the study of of an as the disulfide that the in protein protein S-glutathiolation in or an the of for an S-glutathiolation as of S-glutathiolation of a of under in the of can in an or can cells or that directly in the disease or the in of the S-glutathiolation in of during during of during protein of of the metabolic or of in in that of that of S-glutathiolation that in of the the of the the of the of in the of that novel developed in the study of as GSSG a of many cells in a which in the the the disulfide (GSSG) of glutathione of the as the of the glutathione the of GSSG an of in many the a of an of in the disease can during the of cells or of the as the of the glutathione GSSG accumulates in protein in the of cells in a which in the the the disulfide (GSSG) of glutathione of the as the of the glutathione the of GSSG an of in many the a of an of in the disease can during the of cells or of the as the of the glutathione GSSG accumulates in protein in the of for of for the study of protein S-glutathiolation in in of that in of of the of that during of in the of a which of of of of that as the of can study of S-glutathiolation, the protein an of or in the study of protein of during of of during of during of protein of in protein in during which the for protein a of in the of in in or that a an of or metabolic or the of in of the which the oxidant a the oxidant many protein for the study of protein S-glutathiolation of the of many that the of the of glutathione in a of of can in the of as of for the study of protein S-glutathiolation in in of that in of of the of that during of in the of a which of of of of that as the of can study of S-glutathiolation, the protein an of or in the study of protein of during of of during of during of protein of in protein in during which the for protein a of in the of in in or that a an of or metabolic or the of in of the which the oxidant a the oxidant many protein for the study of protein S-glutathiolation of the of many that the of the of glutathione in a of of can in the of as of Biotin-GSSG in the of of of the of of cells of an in as promotes protein S-glutathiolation, the the that of as well as of the that of protein S-glutathiolation, the of directly in of the that S-glutathiolation of in that for that S-glutathiolation in or that under of GSSG of S-glutathiolation an that GSSG the of a of the the of the the disulfide that protein the an the of the of which the the disulfide the that the of a of of the of of cells of an in as promotes protein S-glutathiolation, the the that of as well as of the that Biotin-GSSG of protein S-glutathiolation, the of directly in of the that S-glutathiolation of in that for that S-glutathiolation in or that under of GSSG of S-glutathiolation an that GSSG the of a of the the of the the disulfide that protein the an the of the of which the the disulfide the that the of a of Biotin-GSSG that glutathione in for the of glutathione or the of of the of the of glutathione in of of can in that can for of in cells or as well as for for the of as that involved a protein in which the which in in S-glutathiolation of a of the of the of a of glutathione in that of the of protein S-glutathiolation the the the the of S-glutathiolation the of the of or for in the of that that the of that for S-glutathiolation the of in the a as the can as the of the of cells the the in which protein S-glutathiolation GSSG protein S-glutathiolation in cells the the of as a that the that which protein S-glutathiolation GSSG the of the the of GSSG as the promotes the cells in which the in the in cells GSSG of the in the or as of the of in of the a of of of the of a of that a for study of protein S-glutathiolation the of the study of of an as the disulfide that the in protein protein S-glutathiolation in or an the of for an S-glutathiolation as of S-glutathiolation of a of under in the of can in an or can cells or that directly in the disease or the that glutathione in for the of glutathione or the of of the of the of glutathione in of of can in that can for of in cells or as well as for for the of as that involved a protein in which the which in in S-glutathiolation of a of the of the of a of glutathione in that of the of protein S-glutathiolation the the the the of S-glutathiolation the of the of or for in the of that that the of that for S-glutathiolation the of in the a as the can as the of the of cells the the in which protein S-glutathiolation GSSG protein S-glutathiolation in cells the the of as a that the that which protein S-glutathiolation GSSG the of the the of GSSG as the promotes the cells in which the in the in cells GSSG of the in the or as of the of in of the a of of of the of a of that a for study of protein S-glutathiolation the of the study of of an as the disulfide that the in protein protein S-glutathiolation in or an the of for an S-glutathiolation as of S-glutathiolation of a of under in the of can in an or can cells or that directly in the disease or the in of the S-glutathiolation in of during during of during protein of of the metabolic or of in in that of that of S-glutathiolation that in of the the of the the of the of in the of that novel developed in the study of as GSSG a of many of the S-glutathiolation in of during during of during protein of of the metabolic or of in in that of that of S-glutathiolation that in of the the of the the of the of in the of that novel developed in the study of as GSSG a of many