title: Sulfation pattern of the fucose branch is important for the anticoagulant and antithrombotic activities of fucosylated chondroitin sulfates.
abstract: BACKGROUND
  The aim is to compare the structures, anticoagulant and antithrombotic activities of two fucosylated chondroitin sulfates isolated from sea cucumbers Isostichopus badionotus (fCS-Ib) and Pearsonothuria graeffei (fCS-Pg), which were known to have different sulfation patterns on the fucose branches.
  
  
  METHODS
  The structures of fCSs were identified using 2D NMR. Anticoagulant activities were measured by activated partial thromboplastin time (APTT) and thrombin time (TT), and inhibition of factors IIa, Xa and XIIa was assessed in vitro. Antithrombotic activity was determined ex vivo by measuring the length and weight of the thrombus generated.
  
  
  RESULTS
  The two fCSs had identical chondroitin sulfate E backbones and similar fucose branches, but different sulfation patterns of the fucose branches. The fucose branch in fCS-Ib was mainly 2,4-O-sulfated whereas that in fCS-Pg was mainly 3,4-O-sulfated. In vitro assay indicated that fCS-Pg possessed much lower potency than fCS-Ib in prolonging APTT/TT and in inhibition of thrombin. However, they both exhibited similar inhibitory effects on factor X activation by intrinsic tenase complex, and on thrombus generation. Furthermore, both fCSs significantly activated factor XII, which has been proved to be associated with adverse clinical events associated with heparin contaminated by oversulfated chondroitin sulfate.
  
  
  CONCLUSIONS
  The 2,4-O-sulfated fucose branch is the key structural factor of fCSs for prolonged APTT/TT and inhibition of thrombin, whereas the inhibitory effect of fCSs on factor X, XII activation and thrombus generation was attributed to the overall structure of fCS polysaccharide. GENERAL IMPORTANCE: Both fCSs have well defined structures and can be readily quality-controlled, and therefore may be potential alternatives for heparin as anticoagulant and antithrombotic drugs.
authors: College of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou 310029, China. chenshiguo210@163.com
authors: Shiguo
authors: S
authors: Chen
authors: Guoyun
authors: G
authors: Li
authors: Nian
authors: N
authors: Wu
authors: Xin
authors: X
authors: Guo
authors: Ningbo
authors: N
authors: Liao
authors: Xingqian
authors: X
authors: Ye
authors: Donghong
authors: D
authors: Liu
authors: Changhu
authors: C
authors: Xue
authors: Wengang
authors: W
authors: Chai
mesh_terms: D000328:Adult; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000494:pharmacology*; D005174:Factor XII / Q000378:metabolism; D065427:Factor Xa Inhibitors; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D012616:Sea Cucumbers / Q000737:chemistry
chemical_list: D000925:Anticoagulants; D065427:Factor Xa Inhibitors; D005343:Fibrinolytic Agents; C517150:fucosylated chondroitin sulfate; D005174:Factor XII; D002809:Chondroitin Sulfates