Chemoenzymatic synthesis and fluorescent visualization of cell-surface selectin-bound sialyl Lewis x derivatives.
Sialyl Lewis x (sLe(x)) derivatives conjugated to readily visualized molecular labels are useful chemical probes to study selectin-carbohydrate interactions. Localization of the selectins on the surface of leukocytes and activated endothelial cells can be detected through fluorescence of bound selectin ligands. Herein we present a short chemoenzymatic synthesis of a fluorescently labeled bivalent sLe(x) conjugate. The use of an amino-substituted monovalent sLe(x) to obtain fluorescent- and biotin-labeled sLe(x) derivatives is also described. The cell-staining utility of the fluorescent sLe(x) conjugates is demonstrated for a HUVEC cell line expressing E-selectin and for CHO-K1 cells expressing either L- or E-selectin.
Wittmann
V
V
Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Datta
A K
AK
Koeller
K M
KM
Wong
C H
CH
D000409:Alanine / Q000737:chemistry; D000818:Animals; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D016466:CHO Cells / Q000378:metabolism; D002240:Carbohydrate Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D005700:Galactosyltransferases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D019041:L-Selectin / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D019087:Selectins / Q000235:genetics / Q000378:metabolism*; D000080506:Sialyl Lewis X Antigen; D013194:Staining and Labeling / Q000379:methods
D019040:E-Selectin; D005456:Fluorescent Dyes; D009844:Oligosaccharides; D019087:Selectins; D000080506:Sialyl Lewis X Antigen; D019041:L-Selectin; D001710:Biotin; D005700:Galactosyltransferases; D000409:Alanine