A novel fluorescent cross-reactive formylpeptide receptor/formylpeptide receptor-like 1 hexapeptide ligand.
Formylpeptide receptors (FPRs) are implicated in a variety of immunological and inflammatory response cascades. Further understanding of FPR-family ligand interactions could play an integral role in biological and therapeutic discovery. Fluorescent reporter ligands for the family are desirable experimental tools for increased understanding of ligand/receptor interactions. The ligand binding affinity and fluorescent reporting activity of the peptide WK(FL)YMVm was explored though use of the high throughput HyperCyt flow cytometric platform. Relative binding affinities of several known FPR and FPRL1 peptide ligands were compared in a duplex assay format. The fluorescent W-peptide ligand, WK(FL)YMVm, proved to be a high-affinity, cross-reactive reporter ligand for the FPR/FPRL1 duplex assay. Ligand specificity was demonstrated for each receptor, with known, selective peptide ligands. The binding site specificity of the reporter ligand was further verified by a fluorescent confocal microscopy internalization experiment. The fluorescent peptide ligand WK(FL)YMVm binds with high affinity to both FPR and FPRL1. The differential affinities of known peptide ligands were observed with the use of this fluorescent probe in high throughput screening flow cytometry.
Strouse
J Jacob
JJ
Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque, 87131, USA.
Young
Susan M
SM
Mitchell
Hugh D
HD
Ye
Richard D
RD
Prossnitz
Eric R
ER
Sklar
Larry A
LA
Edwards
Bruce S
BS
D000818:Animals; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000378:metabolism*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D044042:Receptors, Formyl Peptide / Q000378:metabolism*; D044067:Receptors, Lipoxin / Q000378:metabolism*; D014162:Transfection; D020298:U937 Cells
D002630:Chemotactic Factors; C105101:FPR2 protein, human; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008024:Ligands; D009842:Oligopeptides; D010455:Peptides; D044042:Receptors, Formyl Peptide; D044067:Receptors, Lipoxin; C541378:tryptophyl-(fluoresceinyl-lysyl)-tyrosyl-methionyl-valyl-methioninamide