Large, tissue-regulated domain diversity of heparan sulfates demonstrated by phage display antibodies.
Heparan sulfates (HS) are long, linear polysaccharides with a high degree of variability. They bind to a vast number of proteins such as growth factors and cytokines, and these interactions are likely to be mediated by specific HS domains. To investigate the structural diversity and topological distribution of HS domains in tissues, we selected a panel of 10 unique anti-HS antibodies using phage display technology. All 10 antibodies recognize a specific HS epitope as demonstrated by enzyme-linked immunosorbent assay using defined synthetic HS oligosaccharides, modified HS/heparin molecules, and HS isolated from a variety of organs. The chemical groups involved in the epitopes could be indicated and the position of sulfate groups is of major importance. All HS epitopes have a defined tissue distribution as shown by immunohistochemistry using rat organs. Taken together, the data show that in vivo, a large number of defined HS epitopes exist that do not occur randomly but are tightly, topologically regulated.
Dennissen
Michel A B A
MA
Department of Biochemistry, 194, University Medical Centre, Nijmegen Center for Molecular Life Sciences, University of Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.
Jenniskens
Guido J
GJ
Pieffers
Martijn
M
Versteeg
Elly M M
EM
Petitou
Maurice
M
Veerkamp
Jacques H
JH
van Kuppevelt
Toin H
TH
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics*; D002240:Carbohydrate Sequence; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006497:Heparan Sulfate / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar
D000906:Antibodies; D000939:Epitopes; D006497:Heparan Sulfate