title: Low-molecular-weight heparin and dermatan sulfate end group-labeled with tyramine and fluorescein. Biochemical and biological characterization of the fluorescent-labeled heparin derivative.
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.
authors.affiliation: 1st Department of Medicine, Medical University Clinic, Mannheim, Germany. J-Harenberg@t-online.de
authors.forename: Job
authors.initials: J
authors.lastname: Harenberg
authors.forename: Benito
authors.initials: B
authors.lastname: Casu
authors.forename: Marco
authors.initials: M
authors.lastname: Guerrini
authors.forename: Reinhard
authors.initials: R
authors.lastname: Malsch
authors.forename: Annamaria
authors.initials: A
authors.lastname: Naggi
authors.forename: Lukas
authors.initials: L
authors.lastname: Piazolo
authors.forename: Giangiacomo
authors.initials: G
authors.lastname: Torri
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000493:pharmacokinetics*; D003871:Dermatan Sulfate / Q000737:chemistry / Q000493:pharmacokinetics; D005434:Flow Cytometry; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry / Q000493:pharmacokinetics*; D006493:Heparin / Q000737:chemistry / Q000493:pharmacokinetics; D006495:Heparin, Low-Molecular-Weight / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D014439:Tyramine / Q000737:chemistry
chemical_list: D000925:Anticoagulants; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D006495:Heparin, Low-Molecular-Weight; D003871:Dermatan Sulfate; D006493:Heparin; D019793:Fluorescein; D014439:Tyramine