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dc.rights.licenseOpen Access
dc.contributor.authorToida, Toshihiko
dc.contributor.authorHileman, Ronald E.
dc.contributor.authorSmith, April E.
dc.contributor.authorVlahova, Petinka I.
dc.contributor.authorLinhardt, Robert J.
dc.date1996
dc.date.accessioned2022-06-23T03:52:49Z
dc.date.available2022-06-23T03:52:49Z
dc.date.issued1996-12-30
dc.identifier.citationEnzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites, T. E. Toida, R. E. Hileman, A. E. Smith, P. I. Vlahova, R.J. Linhardt, Journal of Biological Chemistry, 271, 32040-32047, 1996.
dc.identifier.issn219258
dc.identifier.urihttps://hdl.handle.net/20.500.13015/5162
dc.identifier.urihttps://doi.org/10.1074/jbc.271.50.32040
dc.descriptionJournal of Biological Chemistry, 271, 32040-32047
dc.descriptionNote : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
dc.description.abstractTwo new oligosaccharides were prepared from heparin by its partial depolymerization using heparin lyase I (EC 4.2.2.7) in an attempt to prepare oligosaccharides having intact antithrombin III binding sites. The oligosaccharides were purified by chromatography on the basis of both size and charge and demonstrated a high level of purity by capillary electrophoresis. One- and two-dimensional 1H NMR spectroscopy at 500 MHz revealed the structure of each oligosaccharide. The octasaccharide and decasaccharide are ΔUAp2S(1→4)-α-DGlcNpS6S(1→4)-α-L-IdoAp(1→4)-α-D-GlcNpAc6S(1→4)-βD-GlcAp(1→4)-α-D-GlcNpS3S6S(1→4)-α-L-IdoAp2S(1→4)α-D-GlcNpS6S (where ΔUAp is 4-deoxy-α-L-threo-hex-enopyranosyluronic acid, GlcNp is 2-amino-2-deoxy-glucopyranose, GlcAp is glucopyranosyluronic acid, S is sulfate and Ac is acetate) and ΔUAp2S(1→4)-α-D-GlcNpS6S(1→4)-α-L-IdoAp(1→4)-α-D-GlcNpAc6S (1→4)-β-D-GlcAp(1→4)-α-D-GlcNpS3S6S(1→4)-α-L-IdoAp2S(1→4)-α-D-GlcNpS6S(1→4)-α-L-IdoAp2S(1→4)-α-D-GlcNpS6S, respectively. A hexasaccharide containing a similar structural motif to that found in the antithrombin III binding site and having greatly reduced anticoagulant activity was also isolated. The structure of the hexasaccharide is ΔUAp2S(1→4)-α-D-GlcNpAc6S(1→4)-β-D-GlcAp(1→4)-α-D-GlcNpS3S6S(1→4)-α-L-IdoAp(1→4)-α-D-GlcNpS6S. The octasaccharide and decasaccharide correspond to the predominant structural motif found in porcine intestinal mucosal heparin. Sufficient quantities of the decasaccharide were obtained to examine its interaction with antithrombin III using microtitration calorimetry. This decasaccharide bound to antithrombin III with similar avidity as heparin and showed comparable anticoagulant activity, as determined using an antithrombin III dependent anti-factor Xa assay. Interestingly, while both decasaccharide and heparin bound to antithrombin with nanomolar affinity, very little heat of binding was observed.
dc.description.sponsorshipNational Heart, Lung, and Blood Institute
dc.languageen_US
dc.language.isoENG
dc.publisherElsevier
dc.relation.ispartofThe Linhardt Research Labs Online Collection
dc.relation.ispartofRensselaer Polytechnic Institute, Troy, NY
dc.relation.ispartofJournal of Biological Chemistry
dc.relation.urihttps://harc.rpi.edu/
dc.subjectBiology
dc.subjectChemistry and chemical biology
dc.subjectChemical and biological engineering
dc.subjectBiomedical engineering
dc.titleEnzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites
dc.typeArticle
dcterms.accessRightsA full text version is available in DSpace@RPI
dcterms.isPartOfJournal
dcterms.isVersionOfhttps://doi.org/10.1074/jbc.271.50.32040
dc.rights.holderIn Copyright : this Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). https://rightsstatements.org/page/InC/1.0/
dc.creator.identifierhttps://orcid.org/0000-0003-2219-5833
dc.relation.departmentThe Linhardt Research Labs.
dc.relation.departmentThe Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
rpi.description.pages32040-32047
rpi.description.volume271


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