biostudies-literatureHuang YNHLBI NIH HHSNational Heart, Lung, and Blood InstituteNational Institute of General Medical SciencesNIGMS NIH HHS592-600https://www.ebi.ac.uk/biostudies/studies/S-EPMC4287833biostudies-literatureUnknown87(1)Heparan sulfate (HS) 3-O-sulfation determines the binding specificity of HS/heparin for antithrombin III and plays a key role in herpes simplex virus (HSV) infection. However, the low natural abundance of HS 3-O-sulfation poses a serious challenge for functional studies other than the two cases mentioned above. By contrast, multiple distinct isoforms of 3-O-sulfotranserases exist in mammals (up to seven isoenzymes). Here we describe a novel peeling reaction that specifically degrades HS chains with 3-O-sulfated glucosamine at the reducing-end. When HS/heparin is enzymatically depolymerized for compositional analysis, 3-O-sulfated glucosamine at the reducing ends appears to be susceptible to degradation under mildly basic conditions. We propose a 3-O-desulfation initiated peeling reaction mAnalytical chemistryDiscovery of a heparan sulfate 3-O-sulfation specific peeling reaction.PMC4287833P41GM103390R21 HL131554R01HL098950P41 GM103390P41GM104603P41 GM104603Zong CBoons GJHuang YLin CZaia JMao YfalseDiscovery of a heparan sulfate 3-O-sulfation specific peeling reaction.Heparan sulfate (HS) 3-O-sulfation determines the binding specificity of HS/heparin for antithrombin III and plays a key role in herpes simplex virus (HSV) infection. However, the low natural abundance of HS 3-O-sulfation poses a serious challenge for functional studies other than the two cases mentioned above. By contrast, multiple distinct isoforms of 3-O-sulfotranserases exist in mammals (up to seven isoenzymes). Here we describe a novel peeling reaction that specifically degrades HS chains with 3-O-sulfated glucosamine at the reducing-end. When HS/heparin is enzymatically depolymerized for compositional analysis, 3-O-sulfated glucosamine at the reducing ends appears to be susceptible to degradation under mildly basic conditions. We propose a 3-O-desulfation initiated peeling reaction m2015-01-01T00:00:00Z2015 Jan2025-04-04T14:40:18.286Z2019-03-27T01:43:01ZS-EPMC42878332548643710.1021/ac503248k