title: Detection and Removal of Endogenous Carbon Monoxide by Selective and Cell-Permeable Hemoprotein Model Complexes.
abstract: Carbon monoxide (CO) is produced in mammalian cells during heme metabolism and serves as an important signaling messenger. Here we report the bioactive properties of selective CO scavengers, hemoCD1 and its derivative R8-hemoCD1, which have the ability to detect and remove endogenous CO in cells. HemoCD1 is a supramolecular hemoprotein-model complex composed of 5,10,15,20-tetrakis(4-sulfonatophenyl)porphinatoiron(II) and a per-O-methylated β-cyclodextrin dimer having an pyridine linker. We demonstrate that hemoCD1 can be used effectively to quantify endogenous CO in cell lysates by a simple spectrophotometric method. The hemoCD1 assay detected ca. 260 pmol of CO in 106hepatocytes, which was well-correlated with the amount of intracellular bilirubin, the final breakdown product of heme metabolism. We then covalently attached an octaarginine peptide to a maleimide-appended hemoCD1 to synthesize R8-hemoCD1, a cell-permeable CO scavenger. Indeed, R8-hemoCD1 was taken up by intact cells and captured intracellular CO with high efficiency. Moreover, we revealed that removal of endogenous CO by R8-hemoCD1 in cultured macrophages led to a significant increase (ca. 2.5-fold) in reactive oxygen species production and exacerbation of inflammation after challenge with lipopolysaccharide. Thus, R8-hemoCD1 represents a powerful expedient for exploring specific and still unidentified biological functions of CO in cells.
authors.affiliation: Department of Molecular Chemistry and Biochemistry, Faculty of Science and Engineering, Doshisha University , Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Saika
authors.initials: S
authors.lastname: Minegishi
authors.affiliation: Department of Molecular Chemistry and Biochemistry, Faculty of Science and Engineering, Doshisha University , Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Aki
authors.initials: A
authors.lastname: Yumura
authors.affiliation: Department of Molecular Chemistry and Biochemistry, Faculty of Science and Engineering, Doshisha University , Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Hirotsuna
authors.initials: H
authors.lastname: Miyoshi
authors.affiliation: Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts , Kyotanabe, Kyoto 610-0395, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Negi
authors.affiliation: Department of Microbiology, Kansai Medical University , Hirakata, Osaka 573-1010, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Taketani
authors.affiliation: Inserm U955 , Team 12, Créteil 94000, France.|Université Paris Est , Faculty of Medicine, Créteil 94000, France.
authors.forename: Roberto
authors.initials: R
authors.lastname: Motterlini
authors.affiliation: Inserm U955 , Team 12, Créteil 94000, France.|Université Paris Est , Faculty of Medicine, Créteil 94000, France.
authors.forename: Roberta
authors.initials: R
authors.lastname: Foresti
authors.affiliation: Department of Molecular Chemistry and Biochemistry, Faculty of Science and Engineering, Doshisha University , Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Kano
authors.affiliation: Department of Molecular Chemistry and Biochemistry, Faculty of Science and Engineering, Doshisha University , Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Hiroaki
authors.identifier: 0000-0003-4682-7217
authors.initials: H
authors.lastname: Kitagishi
mesh_terms: D000818:Animals; D002248:Carbon Monoxide / Q000032:analysis* / Q000378:metabolism; D002478:Cells, Cultured; D006420:Hemeproteins / Q000737:chemistry* / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D008954:Models, Biological; D015394:Molecular Structure; D000067996:RAW 264.7 Cells
chemical_list: D006420:Hemeproteins; D002248:Carbon Monoxide