| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA
1Correspondence: Massachusetts Institute of Technology, Bldg. 68–480, 77 Massachusetts Ave., Cambridge, MA 02139, USA. E-mail: rdrrosen{at}mit.edu
Heparan sulfate (HS) is a sulfated polysaccharide present on cell surfaces and in the extracellular matrix. Accumulating evidence shows that HS plays key roles in many biological systems by interacting with various proteins in a structural-specific manner. Due to technical difficulties, however, the understanding of critical functional groups on HS for protein interaction is vague. We report a rapid, convenient, sensitive, and inexpensive strategy using in vitro modification with pure enzymes and gel mobility shift assay to study the subject. We demonstrated the requirements of 3-O, 6-O sulfates and the minimal length of oligosaccharide for antithrombin III (AT-III) binding. We regenerated the binding sites for AT-III on completely desulfated N-resulfated heparin and revealed the critical modification enzymes. This new strategy could be used to identify critical functional groups on HS and to generate HS library and biologically active HS, providing information applicable to the design of HS drugs, such as anticoagulant reagents and viral infection blockers. The binding assay with fibroblast growth factors and receptors confirmed the general usefulness of this approach.—Wu, Z. L., Zhang, L., Beeler, D. L., Kuberan, B., Rosenberg, R. D. A new strategy for defining critical functional groups on heparan sulfate.
Key Words: heparin • gel mobility shift assay • in vitro modification • antithrombin III • sulfotransferase
This article has been cited by other articles:
|
|
 |
|
  N. T. Seyfried, C. D. Blundell, A. J. Day, and A. Almond Preparation and application of biologically active fluorescent hyaluronan oligosaccharides Glycobiology, March 1, 2005; 15(3): 303 - 312. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Pisano, C. Aulicino, L. Vesci, B. Casu, A. Naggi, G. Torri, D. Ribatti, M. Belleri, M. Rusnati, and M. Presta Undersulfated, low-molecular-weight glycol-split heparin as an antiangiogenic VEGF antagonist Glycobiology, February 1, 2005; 15(2): 1C - 6C. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Pankonin, J. T. Gallagher, and J. A. Loeb Specific Structural Features of Heparan Sulfate Proteoglycans Potentiate Neuregulin-1 Signaling J. Biol. Chem., January 7, 2005; 280(1): 383 - 388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lyon, J. A. Deakin, D. Lietha, E. Gherardi, and J. T. Gallagher The Interactions of Hepatocyte Growth Factor/Scatter Factor and Its NK1 and NK2 Variants with Glycosaminoglycans Using a Modified Gel Mobility Shift Assay: ELUCIDATION OF THE MINIMAL SIZE OF BINDING AND ACTIVATORY OLIGOSACCHARIDES J. Biol. Chem., October 15, 2004; 279(42): 43560 - 43567. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. K. Powell, E. A. Yates, D. G. Fernig, and J. E. Turnbull Interactions of heparin/heparan sulfate with proteins: Appraisal of structural factors and experimental approaches Glycobiology, April 1, 2004; 14(4): 17R - 30R. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. L. Wu, M. Lech, D. L. Beeler, and R. D. Rosenberg Determining Heparan Sulfate Structure in the Vicinity of Specific Sulfotransferase Recognition Sites by Mass Spectrometry J. Biol. Chem., January 16, 2004; 279(3): 1861 - 1866. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Kuberan, D. L. Beeler, M. Lech, Z. L. Wu, and R. D. Rosenberg Chemoenzymatic Synthesis of Classical and Non-classical Anticoagulant Heparan Sulfate Polysaccharides J. Biol. Chem., December 26, 2003; 278(52): 52613 - 52621. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. L. Wu, L. Zhang, T. Yabe, B. Kuberan, D. L. Beeler, A. Love, and R. D. Rosenberg The Involvement of Heparan Sulfate (HS) in FGF1/HS/FGFR1 Signaling Complex J. Biol. Chem., May 2, 2003; 278(19): 17121 - 17129. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. M. El-Fasakhany, K. Ichihara-Tanaka, K. Uchimura, and T. Muramatsu N-Acetylglucosamine-6-O-Sulfotransferase-1: Production in the Baculovirus System and Its Applications to the Synthesis of a Sulfated Oligosaccharide and to the Modification of Oligosaccharides in Fibrinogen J. Biochem., March 1, 2003; 133(3): 287 - 293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Jemth, J. Kreuger, M. Kusche-Gullberg, L. Sturiale, G. Gimenez-Gallego, and U. Lindahl Biosynthetic Oligosaccharide Libraries for Identification of Protein-binding Heparan Sulfate Motifs. EXPLORING THE STRUCTURAL DIVERSITY BY SCREENING FOR FIBROBLAST GROWTH FACTOR (FGF) 1 AND FGF2 BINDING J. Biol. Chem., August 16, 2002; 277(34): 30567 - 30573. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
