Table 3 Observed first-order rate constants (kobs) for the single turnover conversion of Ac-LKRR-pNa substrate by wild type and mutant forms of both the glycine-linked and unlinked NS2B-NS3pro complexes at 37 °C, pH 8.5, with a 20% excess of enzyme.

From: Product release is rate-limiting for catalytic processing by the Dengue virus protease

 

Glycine-linked

Unlinked

kobs (s−1)

WT

0.82 ± 0.31a

0.88 ± 0.20

L74A

0.69 ± 0.50

1.04 ± 0.70

I76A

0.62 ± 0.31

0.98 ± 0.51

E80A

0.99 ± 0.70

0.70 ± 0.11

S83A

0.78 ± 0.62

0.63 ± 0.22

I86A

0.61 ± 0.30

0.52 ± 0.13

  1. aFor the WT, glycine-linked protease, data was also collected at nearly three- and five-fold excess of enzyme. Corresponding first-order rate constants obtained were 0.79 ± 0.4 s−1 and 0.81 ± 0.3 s−1, respectively, demonstrating that single turnover conditions are attained with a 20% excess of enzyme.
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