Multiparameter flow cytometric analysis of a pH sensitive formyl peptide with application to receptor structure and processing kinetics.
AuthorsFay SP, Habbersett R, Domalewski MD, Posner RG, Houghton TG, Pierson E, Muthukumaraswamy N, Whitaker J, Haugland RP, Freer RJ
JournalCytometry
PubMed ID8168401
'Environmentally sensitive molecules have many potential cellular applications. We have investigated the utility of a pH sensitive ligand for the formyl peptide receptor, CHO-Met-Leu-Phe-Phe-Lys (SNAFL)-OH (SNAFL-seminaphtho-fluorescein), because in previous studies (Fay et al.: Biochemistry 30:5066-5075, 1991) protonation has been used to explain the quenching when the fluoresceinated formyl pentapeptide ligand ... More
Peptide-based targeting of fluorophores to peroxisomes in living cells.
AuthorsPap EH, Dansen TB, Wirtz KW
JournalTrends Cell Biol
PubMed ID11146278
'Peptides carrying different fluorophores can be designed to incorporate spontaneously into living cells when added to the medium. By incorporating the peroxisome-targeting sequence PTS1, the peptide is recognized by the protein-import machinery of peroxisomes and, as a result, can accumulate in these organelles. Depending on the cell type, an inhibitor ... More
Insulin stimulation of retinal outer segment uptake by cultured human retinal pigment epithelial cells determined by a flow cytometric method.
AuthorsMiceli MV, Newsome DA
JournalExp Eye Res
PubMed ID7821371
'This study investigates the mechanism by which insulin stimulates phagocytosis in cultured human retinal pigment epithelium, using a flow cytometric assay of retinal outer segment uptake. RPE cells were isolated from adult human donors and grown in culture. Retinal outer segments were isolated from fresh bovine eyes and covalently labeled ... More
Peroxisomes in human fibroblasts have a basic pH.
AuthorsDansen TB, Wirtz KW, Wanders RJ, Pap EH
JournalNat Cell Biol
PubMed ID10620807
Evaluation of oxidative processes in human pigment epithelial cells associated with retinal outer segment phagocytosis.
AuthorsMiceli MV, Liles MR, Newsome DA
JournalExp Cell Res
PubMed ID8082727
To investigate the nature of the oxidative event that occurs during phagocytosis of retinal outer segments (ROS) by cultured human retinal pigment epithelial (RPE) cells, cells were incubated with isolated bovine ROS labeled with either the fluorescence probe carboxy-SNAFL-2 or the nonfluorescent, oxidizable probe 2',7'-dichlorodihydrofluorescein (H2DCF). The increase in fluorescence ... More
Targeted fluorescent probes in peroxisome function.
AuthorsDansen TB, Pap EHW Wanders RJ, Wirtz KW
JournalHistochem J
PubMed ID11432641
Fluorescent peptides form a new generation of analytical tools for visualizing intracellular processes and molecular interactions at the level of single cells. The peptide-based reporters combine the sensitivity of fluorescence detection with the information specificity of amino acid sequences. Recently we have succeeded in targeting a fluorescent heptapeptide (acetyl-CKGGAKL) carrying ... More