Characterization of anti-heparan sulfate phage display antibodies AO4B08 and HS4E4.
Heparan sulfates (HS) are linear carbohydrate chains, covalently attached to proteins, that occur on essentially all cell surfaces and in extracellular matrices. HS chains show extensive structural heterogeneity and are functionally important during embryogenesis and in homeostasis due to their interactions with various proteins. Phage display antibodies have been developed to probe HS structures, assess the availability of protein-binding sites, and monitor structural changes during development and disease. Here we have characterized two such antibodies, AO4B08 and HS4E4, previously noted for partly differential tissue staining. AO4B08 recognized both HS and heparin, and was found to interact with an ubiquitouys, N-, 2-O-, and 6-O-sulfated saccharide motif, including an internal 2-O-sulfate group. HS4E4 turned out to preferentially recognize low-sulfated HS motifs containing iduronic acid, and N-sulfated as well as N-acetylated glucosamine residues. Contrary to AO4B08, HS4E4 did not bind highly O-sulfated structures such as found in heparin.
Kurup
Sindhulakshmi
S
Department of Medical Biochemistry and Microbiology, University of Uppsala, SE-751 23 Sweden.
Wijnhoven
Tessa J M
TJ
Jenniskens
Guido J
GJ
Kimata
Koji
K
Habuchi
Hiroko
H
Li
Jin-Ping
JP
Lindahl
Ulf
U
van Kuppevelt
Toin H
TH
Spillmann
Dorothe
D
D020816:Amino Acid Motifs; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000939:Epitopes / Q000737:chemistry; D005944:Glucosamine / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry* / Q000276:immunology; D007067:Iduronic Acid / Q000737:chemistry; D019151:Peptide Library; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D000906:Antibodies; D000939:Epitopes; D005944:Glucosamine; D006497:Heparan Sulfate; D007067:Iduronic Acid; D019151:Peptide Library; D011134:Polysaccharides