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Study on complex formation between recombinant human thrombomodulin fragment and thrombin using surface plasmon resonance

✍ Scribed by Kishida, Akio; Nakashima, Mihoko; Sakamoto, Nobuyuki; Serizawa, Takeshi; Maruyama, Ikuro; Akashi, Mitsuru


Book ID
101216128
Publisher
John Wiley and Sons
Year
2000
Tongue
English
Weight
84 KB
Volume
63
Category
Article
ISSN
0361-8609

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✦ Synopsis


Human thrombomodulin (hTM) is a newly described endothelial cell associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. The affinity constant of recombinant human soluble TM (rhs-TM) and a peptide containing active site of hTM fragment (f-hTM) with thrombin were determined using the surface plasmon resonance. The interaction of f-hTM with thrombin could be analyzed by a simple model, whereas the association and the dissociation steps of rhs-TM with thrombin consisted of at least two kinds of interaction phases. The dissociation constant for complex (K D ) of f-hTM and thrombin was determined to be 205 nM, which was more than twice as high as that of rhs-TM (6.7 and 75 nM). The lower affinity of f-hTM was not due to the slow association rate but to the rapid dissociation rate. It comes clear that f-hTM interacts with thrombin rapidly. Am.