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. 2019 Jul 29;9:10914. doi: 10.1038/s41598-019-47494-w

Figure 2.

Figure 2

Kinetic constants for the versicanase activity of ADAMTS-4 and -5. (a) Representative ELISA standard curves of V1 and V1-5GAG digested with ADAMTS-5 (ATS-5). Following complete digestion of either V1 or V1-5GAG (100 nM) with ADAMTS-5, samples were diluted and incubated on a plate coated with anti-DPEAAE antibodies as reported in the Method Section. (b,c) Michaelis-Menten curves for proteolysis of V1-5GAG by ADAMTS-4 (b) and -5 (c). The enzyme (5.5 nM ADAMTS-4, 0.2 nM ADAMTS-5) was incubated with increasing concentrations of substrate. At the indicated time points, an aliquot was taken, proteolysis stopped with EDTA and cleavage products measured by ELISA. Data are plotted as turnover number versus substrate concentration and are presented as mean ± SEM (n = 3–4).