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Semin Thromb Hemost 2002; 28(4): 343-354
DOI: 10.1055/s-2002-34303
Copyright © 2002 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA. Tel.: +1(212) 584-4662

Low-Molecular-Weight Heparin and Dermatan Sulfate End Group-Labeled with Tyramine and Fluorescein. Biochemical and Biological Characterization of the Fluorescent-Labeled Heparin Derivative

Job Harenberg1 , Benito Casu2 , Marco Guerrini2 , Reinhard Malsch1 , Annamaria Naggi2 , Lukas Piazolo1 , Giangiacomo Torri2
  • 11st Department of Medicine, Medical University Clinic, Mannheim, Germany
  • 2Istituto di Chimica e Biochimica "G. Ronzoni", Milan, Italy
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Publication History

Publication Date:
23 September 2002 (online)

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ABSTRACT

To improve the understanding of the biological functions and pharmacology of heparin and dermatan sulfate, low-molecular-weight heparin (LMWH) and low-molecular-weight dermatan sulfate (LMWDS) were labeled with tyramine (T) by covalently linking T to the terminal residue of 2,5-anhydromannose (or 2,5-anhydrotalose for dermatan sulfate). The covalent labeling was demonstrated by nuclear magnetic resonance spectroscopy. The tyramine-labeled LMWH (LMWH-T) was also labeled with fluorescein (F) by further reacting it with fluorescein isothiocyanate. The fluoresceinated LMWH-T (LMWH-T,F ) was used to analyze biological functions on blood coagulation and binding to leukocytes. The biological activities on factor Xa and thrombin inhibition remained unchanged compared with the parent compound. Flow cytometric analysis of leukocytes demonstrated binding of the modified heparin to granulocytes, monocytes, and lymphocytes, the half-live being twice as long as the antifactor Xa activity. F-labeled heparin was displaced by unlabeled heparin from all three populations of leukocytes. Binding of heparin to leukocytes may play an important role in inflammation and atherosclerosis.

KEYWORDS

Glycosaminoglycans - heparin - dermatan sulfate - tyramine- and fluorescent-labeled low-molecular-weight heparin - leukocytes

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