Table 1

Specificity constants for ADAMTS13 variants and VWF substrates

SubstrateADAMTS13 construct, kcat/Km × 104 M−1s−1
MDMDTMDTCMDTCSFL
VWF73 0.61 ± 0.02 0.80 ± 0.05 8.1 ± 0.5 205 ± 66* 75 ± 20* 
VWF64 0.61 ± 0.04 0.95 ± 0.02 6.4 ± 0.2 10 ± 3* 5 ± 3* 
VWF46 0.022 ± 0.001 0.29 ± 0.02 0.90 ± 0.04 0.98 ± 0.02 1.02 ± 0.06 
VWF35 0.044 ± 0.004 0.34 ± 0.03 0.90 ± 0.05 1.12 ± 0.02 1.2 ± 0.1 
VWF28 0.047 ± 0.002 0.06 ± 0.004 0.17 ± 0.09 0.21 ± 0.01 0.2 ± 0.01 
VWFd1 0.58 ± 0.02 0.7 ± 0.1 1.1 ± 0.1  2.4 ± 0.7 
VWFd2 0.46 ± 0.03 0.9 ± 0.1 2.6 ± 0.2  8.4 ± 1.7 
VWFd3 0.81 ± 0.04 0.76 ± 0.04 10 ± 0.3  25 ± 5.5 
SubstrateADAMTS13 construct, kcat/Km × 104 M−1s−1
MDMDTMDTCMDTCSFL
VWF73 0.61 ± 0.02 0.80 ± 0.05 8.1 ± 0.5 205 ± 66* 75 ± 20* 
VWF64 0.61 ± 0.04 0.95 ± 0.02 6.4 ± 0.2 10 ± 3* 5 ± 3* 
VWF46 0.022 ± 0.001 0.29 ± 0.02 0.90 ± 0.04 0.98 ± 0.02 1.02 ± 0.06 
VWF35 0.044 ± 0.004 0.34 ± 0.03 0.90 ± 0.05 1.12 ± 0.02 1.2 ± 0.1 
VWF28 0.047 ± 0.002 0.06 ± 0.004 0.17 ± 0.09 0.21 ± 0.01 0.2 ± 0.01 
VWFd1 0.58 ± 0.02 0.7 ± 0.1 1.1 ± 0.1  2.4 ± 0.7 
VWFd2 0.46 ± 0.03 0.9 ± 0.1 2.6 ± 0.2  8.4 ± 1.7 
VWFd3 0.81 ± 0.04 0.76 ± 0.04 10 ± 0.3  25 ± 5.5 

Reactions were performed at room temperature and products were analyzed by MALDI-TOF MS. ADAMTS13 construct M did not cleave any substrate, and no other ADAMTS13 construct cleaved substrate VWFd4 or VWFd5. Values (± SD) are based on 4 to 12 independent rate determinations.

*

Data from Gao et al are provided for comparison.17 

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