Intracellular oxygen-sensitive phosphorescent probes based on cell-penetrating peptides.
Probing of molecular oxygen in mammalian cells is important for the analysis of mitochondrial function, metabolic responses, and energetic status of the cells. We describe a new panel of intracellular O(2)-sensitive probes based on phosphorescent porphyrin dyes conjugated to cell-penetrating peptides. The probes comprising the uncharged derivatives of Pt(II)-coproporphyrin I covalently linked to positively charged TAT-derived peptides are shown to effectively load live mammalian cells without any transfection reagents. The probes work well with all cell types tested, show similar subcellular localization, and produce characteristic responses to cell stimulation with mitochondrial uncouplers and inhibitors. They provide a simple and versatile tool for O(2) monitoring in live cells and in tissue, and an alternative to the existing O(2) probes which require facilitated transport into the cell.
Dmitriev
Ruslan I
RI
Biochemistry Department, University College Cork, Cavanagh Pharmacy Building, Cork, Ireland.
Zhdanov
Alexander V
AV
Ponomarev
Gelii V
GV
Yashunski
Dmitri V
DV
Papkovsky
Dmitri B
DB
D000595:Amino Acid Sequence; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry*; D045325:HCT116 Cells; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D010100:Oxygen / Q000032:analysis*; D016716:PC12 Cells; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010984:Platinum / Q000737:chemistry; D011166:Porphyrins / Q000737:chemistry*; D051381:Rats
D005456:Fluorescent Dyes; D010455:Peptides; D011166:Porphyrins; D010984:Platinum; D010100:Oxygen