RRC ID 66051
Author Minegishi S, Yumura A, Miyoshi H, Negi S, Taketani S, Motterlini R, Foresti R, Kano K, Kitagishi H.
Title Detection and Removal of Endogenous Carbon Monoxide by Selective and Cell-Permeable Hemoprotein Model Complexes.
Journal J Am Chem Soc
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 106 hepatocytes, 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.
Volume 139(16)
Pages 5984-5991
Published 2017-4-26
DOI 10.1021/jacs.7b02229
PMID 28388069
MeSH Animals Carbon Monoxide / analysis* Carbon Monoxide / metabolism Cells, Cultured Hemeproteins / chemistry* Hemeproteins / metabolism Hep G2 Cells Humans Mice Microscopy, Confocal Models, Biological* Molecular Structure RAW 264.7 Cells
IF 14.695
Human and Animal Cells Hep G2(RCB1886) HeLa(RCB0007) RAW 264(RCB0535) NIH3T3(RCB1862)