Enzymatic modification of heparan sulfate on a biochip promotes its interaction with antithrombin III.
A heparan sulfate glycosaminoglycan chain, biotinylated at its reducing-end, was bound to a streptavidin-coated biochip. Surface plasmon resonance spectroscopy showed a low affinity interaction with antithrombin III (ATIII) when it was flowed over a surface containing heparan sulfate. ATIII bound tightly with high affinity when the same surface was enzymatically modified to using 3-O-sulfotransferase isoform 1 (3-OST-1) in the presence of 3'-phosphoadenosine 5'-phosphosulfate (PAPS). The 3-OST-1 enzyme is involved in heparan sulfate biosynthesis and introduces a critical 3-O-sulfo group into this glycosaminoglycan affording the appropriate pentasaccharide sequence capable of high affinity binding to ATIII. This experiment demonstrates the specific structural modification of a glycosaminoglycan bound to a biochip using a biosynthetic enzyme, suggesting a new approach to rapid screening glycosaminoglycan-protein interactions.
Hernaiz
M
M
Division of Medicinal and Natural Products Chemistry, University of Iowa, Iowa City, Iowa, 52242, USA.
Liu
J
J
Rosenberg
R D
RD
Linhardt
R J
RJ
D000818:Animals; D000990:Antithrombin III / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D006497:Heparan Sulfate / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D009844:Oligosaccharides; D011485:Protein Binding
D000990:Antithrombin III; D006497:Heparan Sulfate; D009844:Oligosaccharides