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Nature of Interaction of Marine Sulfated Polysaccharide-911 with V3 Region of gp120 Analysis by a Novel Method of Immobilization of 911 for Surface plasmon Resonance(SPR) Biosensor

Nature of Interaction of Marine Sulfated Polysaccharide-911 ...

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2002全国青年药理学英文学术会议

作者： LI Fu-chuan, GENG Mei-yu, QI Xin Marine Drug & Food Institute, Ocean University of Qingdao, china, 266003

AIM: Surface plasmon resonance (SPR) biosensor was used to monitor the interaction of marine sulfated polysaccharide-911with V3 region of HIV-1 MN gp120, employing 911 immobilized on a sensor chip surface. METHODS: A method for the immobilization of polysaccharides to the biosensor chip surface is described to avoid impairing the internal structure of 911, which was simple and applicable to the immobilization of many reductive polysaccharides and oligosaccharides. Primary amine group was introduced to the reductive end of 911 by reductive amination reaction with 1,6-hexanediamine and was biotinylated further via the reaction of the primary amine with sulfo-NHS-biotin. The biotinylated 911 at reductive end was immobilized on the surface of biosensor chip CM5 by streptavidin-biotin coupling. The interaction of 911 with V3 region was discussed. The results indicated that about 3 molecules of V3 region can bind to a 911 chain and the dissociation constants are estimated to be approximately 10-100nM. CONCLUSION: It is the first time for us to elucidate the molecule interaction of 911 with V3 region of gp120, giving us a better understanding of the mechanism of actions underlying Anti-HIV activity of 911 at its molecule level.

关键词： polysacchande reductive animation SPR V3 region

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Study on Fluorescent Labeling of Marine Sulfated Polysaccharide 911

Study on Fluorescent Labeling of Marine Sulfated Polysacchar...

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2002全国青年药理学英文学术会议

作者： LI Fu-chuan, FAN Ying, LI Jing, GENG Mei-yu (Institute of Marine Drug and Food, Ocean University of Qingdao, Qingdao 266003)

The reducing terminal of marine sulfated polysaccharide (911) was used to selectively insert primary and secondary amines by reductive amination. 911 was bound to 4-(2-aminoethylphenol), followed by labeling the 911-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. High-performance size-exclusion chromatography (HPSEC), ultraviolet/visible (UV/Vis) spectroscopy and nuclear magnetic resonance spectroscopy (NMR) demonstrated the binding of tyramine to 911 and the labeling of FITC to 911-Tyr respectively. Furthermore, H-NMR spectra and UV/Vis spectra revealed about 60％ binding of tyramine to 911 and about 80％ labeling of FITC to 911-Tyr. The anticoagulant activity of labeled fluorescent compound exhibited 15u/mg equivalent to that of unlabled 911. In addition, labeled compound demonstrated to fluorescently stain lymphocytes without any cytotoxity. Taken together, the selectively "end point attached" method might be widely used for fluorecent labeling to those polysaccharides with reducing terminal.

关键词： sulfated polysacchande reductive animation fluorescent labeling

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