Author(s)

Jian Tang¹, Wei Huang¹, Ning Shen¹, Tao Hong², Suying Dang^3*, Wei Zhang^1,2,4*

¹Department of Pharmacology, Institutes for Advanced Interdisciplinary Research, East China Normal University, Shanghai, China.
²Key Laboratory of Brain Functional Genomics (East China Normal University), Ministry of Education, Shanghai, China.
³Department of Biochemistry and Molecular Cell Biology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁴Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, Shanghai, China.

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.

ADAMTS-18, Thrombin, Cleavage, Mass Spectrum

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.