Quantitation of dermatan sulfate active site for heparin cofactor II by 1H nuclear magnetic resonance spectroscopy.
The sequence (IdoA2SO3-GalNAc4SO3)n contributes to the HCII-mediated inhibition of thrombin by dermatan sulfate (DS). This sequence clearly results from the 13C NMR spectrum and can be quantified by the signal C1-H of IdoA2SO3 in the 1H NMR spectrum. A linear correlation has been found between the content in the disulfated disaccharide delta Di-2,4diS obtained by enzymatic demolition with ABC lyase, the percentage content in IdoA2SO3 quantified by 1H NMR, and the HCII-mediated activity of dermatan sulfates from beef mucosa and pig skin. DSs have been obtained also from pig mucosa and contain an amount, not negligible, of delta Di-4, 6diS. This disulfate disaccharide contributes to the activity expressed by the IdoA2SO3-GalNAc4SO3 sequence. The analytical techniques HPLC and 1H NMR, applied to the currently performed analyses of DS, are described and discussed.
Mascellani
G
G
Opocrin S.p.A. R&D Laboratorio, Corlo di Formigine, Modena, Italy.
Liverani
L
L
Prete
A
A
Bergonzini
G
G
Bianchini
P
P
Torri
G
G
Bisio
A
A
Guerrini
M
M
Casu
B
B
D001665:Binding Sites; D003871:Dermatan Sulfate / Q000032:analysis* / Q000378:metabolism; D015844:Heparin Cofactor II / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy
D003871:Dermatan Sulfate; D015844:Heparin Cofactor II