caa a22 4500 605507732 CHVBK 20210128100633.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00775-015-1265-3 doi (NATIONALLICENCE)springer-10.1007/s00775-015-1265-3 An effective method for profiling the selenium-binding proteins using its reactive metabolic intermediate [Elektronische Daten] [Eriko Hori, Sakura Yoshida, Mamoru Haratake, Sakiko Ura, Takeshi Fuchigami, Morio Nakayama] Currently, the intracellular reduction and/or transport of selenium still remain unknown. Certain reduced forms of selenium species are thought to be reactive with various endogenous molecules, particularly thiol-containing proteins. In this study, a profiling method for identifying the selenium-binding proteins using l-penicillamine selenotrisulfide (PenSSeSPen) as a model of the selenium metabolic intermediate was applied to the cell lysate generated from the rat liver. Several proteins with cysteine thiol were found to be reactive with PenSSeSPen through the thiol-exchange reaction by MALDI TOF-MS analysis. The most distinctive cysteine-containing protein at m/z 14,313 in the liver cell lysate was identified as the liver fatty acid-binding protein based on a rat protein database search and a tryptic fragmentation experiment. This methodology could be used for determining the selenium-binding proteins and/or selenium-interactive species and provide a better understanding of the selenium metabolism and utilization in biological systems. SBIC, 2015 Liver fatty acid-binding protein nationallicence Mass spectrometry nationallicence Selenium nationallicence Selenotrisulfide nationallicence Thiol exchange nationallicence Cys : l-Cysteine nationallicence DAN : 2,3-Diaminonaphthalene nationallicence DTNB : 5,5′-Dithiobis(2-nitrobenzoic acid) nationallicence GPx : Glutathione peroxidase nationallicence GSH : Glutathione in the reduced form nationallicence LFABP : Liver fatty acid-binding protein nationallicence MALDI TOF-MS : Matrix-assisted laser desorption ionization time of flight-mass spectrometry nationallicence NEM : N-Ethylmaleimide nationallicence Pen : L-Penicillamine nationallicence PenSSeSPen : L-Penicillamine selenotrisulfide nationallicence SeCys, Sec and U : l-Selenocysteine nationallicence STS : Selenotrisulfide nationallicence Hori Eriko Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut Yoshida Sakura Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut Haratake Mamoru Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, 860-0082, Kumamoto, Japan aut Ura Sakiko Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut Fuchigami Takeshi Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut Nakayama Morio Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/5(2015-07-01), 781-789 0949-8257 20:5<781 2015 20 775 https://doi.org/10.1007/s00775-015-1265-3 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00775-015-1265-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hori Eriko Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Yoshida Sakura Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Haratake Mamoru Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Ura Sakiko Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Fuchigami Takeshi Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Nakayama Morio Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 773 0- JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/5(2015-07-01), 781-789 0949-8257 20:5<781 2015 20 775