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• W2046037847 endingPage "170" @default.
• W2046037847 startingPage "162" @default.
• W2046037847 abstract "Thrombomodulin (TM) on endothelial cells is a glycoprotein that functions as a cofactor for thrombin-catalyzed activation of protein C. The structural requirement for thrombin binding and cofactor activity were investigated using monoclonal antibodies (moAbs) against TM and site-directed mutagenesis of recombinant human soluble TM (rsTM). Results showed that moAb 2A2 inhibited thrombin binding to rsTM and also abolished its functions as a cofactor in thrombin-catalyzed activation of protein C and as an anticoagulant by modifying thrombin-induced fibrinogen clotting and platelet aggregation, moAb 1F2 did not affect its activity as an anticoagulant, but inhibited its cofactor activity, and moAb 10A3 did not inhibit either activity. Epitope analysis was carried out by site directed mutagenesis of rsTM expressed in CHO cells. Some proteins with mutations within the second disulfide loop of the fourth EGF-like domain showed reduced affinity for moAb 1F2, but retained cofactor activity. These results suggest that the epitope of moAb 1F2 includes the second disulfide loop of the fourth EGF-like domain, which is close to a region required for cofactor activity. Mutant proteins of the third disulfide loop of the fifth EGF-like domain showed loss of interaction with moAb 2A2. Thus the epitope of moAb 2A2 may include the third disulfide loop of the fifth EGF-like domain. Furthermore, replacement of Asn-439 by Gln decreased the cofactor activity and anticoagulant activity, and resulted in low affinity for either moAb 1F2 or 2A2, suggesting that Asn-439, which is located in the second disulfide loop of the sixth EGF-like domain, is critical for determining the functional conformation of the EGF-like domains 4–6." @default.
• W2046037847 created "2016-06-24" @default.
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• W2046037847 date "1994-04-01" @default.
• W2046037847 modified "2023-09-26" @default.
• W2046037847 title "Monoclonal antibodies against human thrombomodulin whose epitope is located in epidermal growth factor-like domains" @default.
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• W2046037847 doi "https://doi.org/10.1016/0167-4838(94)90229-1" @default.
• W2046037847 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/7512385" @default.
• W2046037847 hasPublicationYear "1994" @default.
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