Structural Determinants Of Heparan Sulfate Interactions With Slit Proteins

Zhang, Fuming
Ronca, Francesca
Linhardt, Robert J.
Margolis, Richard U.
No Thumbnail Available
Other Contributors
Issue Date
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Terms of Use
In Copyright : this Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
Full Citation
Structural Determinants Of Heparan Sulfate Interactions With Slit Proteins, F. Zhang, F. Ronca, R.J. Linhardt, R.U. Margolis, Biochemical Biophysical Research Communications, 317, 352-357, 2004.
We have previously demonstrated that the Slit proteins, which are involved in axonal guidance and related processes, are high-affinity ligands of the heparan sulfate proteoglycan glypican-1. Glypican–Slit protein interactions have now been characterized in greater detail using two approaches. The ability of heparin oligosaccharides of defined structure (ranging in size from disaccharide to tetradeccasaccharide) to inhibit binding of a glypican-Fc fusion protein to recombinant human Slit-2 was determined using an ELISA. Surface plasmon resonance (SPR) spectroscopy, which measures the interactions in real time, was applied for quantitative modeling of heparin–Slit binding on heparin biochips. Heparin was covalently immobilized on these chips through a pre-formed albumin–heparin conjugate, and the inhibition of Slit binding by heparin, LMW heparin, and heparin-derived oligosaccharides (di-, tetra-, hexa-, and octa-) was examined utilizing solution competition SPR. These competition studies demonstrate that the smallest heparin oligosaccharide competing with heparin binding to Slit was a tetrasaccharide, and that in the ELISA maximum inhibition (∼60% at 2 μM concentration) was attained with a dodecasaccharide.
Biochemical Biophysical Research Communications, 317, 352-357
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Biochemical and Biophysical Research Communications