Enzymatic redesigning of biologically active heparan sulfate.
Heparan sulfate carries a wide range of biological activities, regulating blood coagulation, cell differentiation, and inflammatory responses. The sulfation patterns of the polysaccharide are essential for the biological activities. In this study, we report an enzymatic method for the sulfation of multimilligram amounts of heparan sulfate with specific functions using immobilized sulfotransferases combined with a 3'-phosphoadenosine 5'-phosphosulfate regeneration system. By selecting appropriate enzymatic modification steps, an inactive precursor has been converted to the heparan sulfate having three distinct biological activities, associated with binding to antithrombin, fibroblast growth factor-2, and herpes simplex virus envelope glycoprotein D. Because the recombinant sulfotransferases are expressed in bacteria, and the method uses a low cost sulfo donor, it can be readily utilized to synthesize large quantities of anticoagulant heparin drug or other biologically active heparan sulfates.
Chen
Jinghua
J
Division of Medicinal Chemistry and Natural Products, School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Avci
Fikri Y
FY
Muñoz
Eva M
EM
McDowell
Lynda M
LM
Chen
Miao
M
Pedersen
Lars C
LC
Zhang
Lijuan
L
Linhardt
Robert J
RJ
Liu
Jian
J
D000818:Animals; D000925:Anticoagulants / Q000737:chemistry; D000991:Antithrombins / Q000737:chemistry; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004187:Disaccharides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D015951:Factor Xa / Q000737:chemistry; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D006493:Heparin / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry*; D007249:Inflammation; D020128:Inhibitory Concentration 50; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010724:Phosphoadenosine Phosphosulfate / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D018139:Simplexvirus / Q000235:genetics; D015238:Sulfotransferases / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013917:Thrombin / Q000737:chemistry; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000737:chemistry
D000925:Anticoagulants; D000991:Antithrombins; D004187:Disaccharides; D015951:Factor Xa; D016222:Fibroblast Growth Factor 2; D006493:Heparin; D006497:Heparan Sulfate; D010724:Phosphoadenosine Phosphosulfate; D011134:Polysaccharides; D011994:Recombinant Proteins; D015238:Sulfotransferases; D013917:Thrombin; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1