Table S3.
Steady-state kinetics of AtGH3.5 acyl binding site mutants
| Protein | kcat IAA, min−1 | KmIAA, μM | kcat/KmIAA, M−1⋅s−1 | kcat BA, min−1 | KmBA, μM | kcat/KmBA, M−1⋅s−1 |
| AtGH3.5 | 14.5 ± 1.4 | 770 ± 110 | 314 | 16.0 ± 0.8 | 790 ± 60 | 338 |
| Y134W | 3.7 ± 0.3 | 3,060 ± 310 | 20 | 7.7 ± 0.4 | 6,680 ± 700 | 19 |
| V174L | 7.2 ± 2.8 | 6,720 ± 330 | 18 | 3.1 ± 0.3 | 7,910 ± 1,250 | 7 |
| V231L | 22.0 ± 5.4 | 4,380 ± 126 | 84 | 9.4 ± 0.3 | 4,120 ± 270 | 38 |
| M337L | 13.5 ± 1.5 | 2,930 ± 470 | 77 | 100 ± 47 | 5,650 ± 310 | 295 |
| A339V | 18.0 ± 5.1 | 3,770 ± 130 | 80 | 21.0 ± 0.7 | 1,320 ± 90 | 265 |
| Y344W | 4.8 ± 1.3 | 3,600 ± 100 | 21 | 6.1 ± 0.6 | 4,550 ± 620 | 22 |
AtGH3.5 values are from Table S2 and are shown for comparison with the mutant enzymes. Average values ± SD (n = 3–5) are shown for steady-state kinetic parameters. Assays were performed as described with fixed concentrations of ATP (1 mM) and Asp (5 mM) and with varied concentrations of IAA and BA.