title: Direct determination of the redox status of cysteine residues in proteins in vivo.
abstract: The redox states of proteins in cells are key factors in many cellular processes. To determine the redox status of cysteinyl thiol groups in proteins in vivo, we developed a new maleimide reagent, a photocleavable maleimide-conjugated single stranded DNA (DNA-PCMal). The DNA moiety of DNA-PCMal is easily removed by UV-irradiation, allowing DNA-PCMal to be used in Western blotting applications. Thereby the state of thiol groups in intracellular proteins can be directly evaluated. This new maleimide compound can provide information concerning redox proteins in vivo, which is important for our understanding of redox networks in the cell.
authors.affiliation: Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-ku, Yokohama 226-8503, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hara
authors.affiliation: Dojindo Laboratories, 2025-5 Tabaru, Mashiki-machi, Kumamoto 861-2202, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tatenaka
authors.affiliation: Dojindo Laboratories, 2025-5 Tabaru, Mashiki-machi, Kumamoto 861-2202, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Ohuchi
authors.affiliation: Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-ku, Yokohama 226-8503, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo 102-0075, Japan. Electronic address: thisabor@res.titech.ac.jp.
authors.forename: Toru
authors.initials: T
authors.lastname: Hisabori
mesh_terms: D017360:Arabidopsis / Q000201:enzymology; D015153:Blotting, Western; D003545:Cysteine / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D008301:Maleimides / Q000737:chemistry*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011506:Proteins / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D008301:Maleimides; D011506:Proteins; D013438:Sulfhydryl Compounds; C043592:maleimide; D004247:DNA; D003545:Cysteine
keywords: Cysteine; Photocleavable maleimide-conjugated DNA; Sulfhydryl; Thiol reagent; Thiol redox status; Western blotting