title: A Set of Organelle-Localizable Reactive Molecules for Mitochondrial Chemical Proteomics in Living Cells and Brain Tissues.
abstract: Protein functions are tightly regulated by their subcellular localization in live cells, and quantitative evaluation of dynamically altered proteomes in each organelle should provide valuable information. Here, we describe a novel method for organelle-focused chemical proteomics using spatially limited reactions. In this work, mitochondria-localizable reactive molecules (MRMs) were designed that penetrate biomembranes and spontaneously concentrate in mitochondria, where protein labeling is facilitated by the condensation effect. The combination of this selective labeling and liquid chromatography-mass spectrometry (LC-MS) based proteomics technology facilitated identification of mitochondrial proteomes and the profile of the intrinsic reactivity of amino acids tethered to proteins expressed in live cultured cells, primary neurons and brain slices. Furthermore, quantitative profiling of mitochondrial proteins whose expression levels change significantly during an oxidant-induced apoptotic process was performed by combination of this MRMs-based method with a standard quantitative MS technique (SILAC: stable isotope labeling by amino acids in cell culture). The use of a set of MRMs represents a powerful tool for chemical proteomics to elucidate mitochondria-associated biological events and diseases.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University , Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yasueda
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University , Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Tamura
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University , Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
authors.forename: Alma
authors.initials: A
authors.lastname: Fujisawa
authors.affiliation: Institute of Transformative Bio-Molecules (ITbM), Nagoya University , Chikusa, Nagoya, 464-8602, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kuwata
authors.affiliation: Frontier Research Institute for Materials Science (FRIMS), Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan.|Department of Life Science and Applied Chemistry, Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Tsukiji
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University , Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kiyonaka
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University , Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.|CREST (Core Research for Evolutional Science and Technology, JST) , Sanbancho, Chiyodaku, Tokyo, 102-0075, Japan.
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D018929:Cell Culture Techniques; D002851:Chromatography, High Pressure Liquid; D006367:HeLa Cells; D006801:Humans; D007553:Isotope Labeling; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry*; D008958:Models, Molecular; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012235:Rhodamines / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D024101:Mitochondrial Proteins; D020543:Proteome; D012235:Rhodamines; C029773:rhodamine B