Synthesis of a fluorescent ganglioside GM1 derivative and screening of a synthetic peptide library for GM1 binding sequence motifs.
A ganglioside GM1 probe bearing a dark-red fluorescent dye at the sphingosine moiety of the molecule was prepared by a convenient one-pot synthesis. The labeled GM1 permitted the detection of the natural ganglioside GM1 ligand Escherichia coli heat-labile enterotoxin subunit B (EtxB) in picomole quantities on a solid support. When an epitope mapping of several ganglioside binding proteins and protein fragments was performed by screening a cellulose membrane-bound synthetic library of 64 16mer peptides with the new probe, several peptides displaying ganglioside GM1 affinity could be identified. We consider the labeled glycolipid described herein a versatile tool for manifold biochemical investigations.
Röckendorf
Niels
N
Division of Mucosal Immunology, Department of Clinical Medicine, Research Center Borstel, Borstel, Germany.
Bade
Steffen
S
Hirst
Timothy R
TR
Gorris
Hans H
HH
Frey
Andreas
A
D020816:Amino Acid Motifs; D001427:Bacterial Toxins / Q000737:chemistry*; D001665:Binding Sites; D004768:Enterotoxins / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry*; D005453:Fluorescence; D005677:G(M1) Ganglioside / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D019151:Peptide Library
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D008024:Ligands; D010446:Peptide Fragments; D019151:Peptide Library; D005677:G(M1) Ganglioside; C027538:heat-labile enterotoxin, E coli