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Chondroitin Sulfate in the Preparation of Glycosaminoglycan from Mackerel(Pneumatophorus japonicus Houttuyn) Nasal Cartilage and Its Interaction with Growth Factors |
Yue-Gang ZHOU |
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Abstract To search for new resource of chondroitin sulfate (CS) for therapeutics, glycosaminoglycan (GAG) was isolated from nasal proteoglycan(PG) of mackerel (Pneumatophorus japonicus Houttuyn). The disaccharide composition of CS in the GAG preparation was determined by anion-exchange HPLC after digestion with chondroitinase. The molecular size of GAG was determined by gel filtration, and the interaction of GAG with growth factors was analyzed to determine the kinetic parameter ka(association rate constant)、 kd (dissociation rate constant) and KD(equilibrium dissociation constant) through surface plasmon resonance (SPR) analysis. The content of GAG was 1.09 μmol/mg PG(as disaccharide unites) or 651 μg/mg PG. CS is the major ingredient of the GAG preparation (1.03 μmol/mg PG). Unsaturated disaccharide units produced from CS were ΔDi-6S(38.8%), ΔDi-4S(46.3%), ΔDi-0S(8.4%) and ΔDi-diSD(6.5%). Molecular size of GAG (CS) was approximately 78 kD. The kinetic parameter ka((mol/L)-1·s-1), kd(s-1) and kD(nmol/L) of GAG (CS) with the growth factors were (2.77±0.17)×105, (7.74±1.56)×10-5 and (0.28±0.06) for midkine (MK); (1.05±0.22)×104, (4.16±0.80)×10-3 and (417±131.3) for pleiotrophin (PTN); and (7.04±0.94)×105, (7.84±2.82)×10-3 and (11.1±3.80) for hepatocyte growth factor (HGF), respectively. High affinity with MK and HGF and higher affinity with PTN was observed, suggesting that the CS chains in mackerel nasal cartilage may regulate growth factor signaling and have a potential for therapeutic applications to some diseases
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Received: 23 June 2011
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Corresponding Authors:
Yue-Gang ZHOU
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