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Arg-Ser-775, 792 and 823 in Spacer Region of ADAMTS-18 Is Critical for Thrombin Cleavage

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Cleavage of ADAMTS-18 by thrombin represents a new mechanism of platelet thrombus clearance via the release of active ~45-kDa C-terminal fragments that induces oxidative platelet fragmentation. The exact cleavage sites remain unclear, but Arg (R)775/Ser (S)776 in spacer region of ADAMTS-18 has been shown to be one of the cleavage sites of thrombin. Here, we demonstrate that R792/S793 and R823/S824 are also thrombin cleavage sites by sequence analysis, amino acid mutation and mass spectrometry assay. All these cleavage sites are thrombin-specific and insensitive to other enzymes tested (e.g. cathepsin D or trypsin). Simultaneous mutation of R775, 792, 823 to S775, 792, 823 in ADAMTS-18 completely abrogated the cleavage by thrombin and the generation of active C-terminal 45-kDa fragments. Together with previous study, a total of three thrombin-specific cleavage sites have been identified in spacer region of ADAMTS-18.
Cite this paper
Tang, J. , Huang, W. , Shen, N. , Hong, T. , Dang, S. and Zhang, W. (2014) Arg-Ser-775, 792 and 823 in Spacer Region of ADAMTS-18 Is Critical for Thrombin Cleavage. Health, 6, 2490-2498. doi: 10.4236/health.2014.618286. 
 

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