TABLE 1.
Steady-state kinetic parameters for the reduction of AGE precursors by AKRs
| Substrates | Genbank accession number | Proteins | Km (μM) | kcat (min−1) | kcat/Km (min−1/μM−1) |
|---|---|---|---|---|---|
| Glyoxal | NM_021473 | AKR1A4 | 3,229 ± 904 | 45.8 ± 4.2 | 0.014 |
| NM_001628 | AKR1B1 | 350 ± 20 | 29.5 ± 2.02 | 0.082 | |
| NM_009658 | AKR1B3 | 333 ± 62 | 12.1 ± 0.36 | 0.036 | |
| NM_009731 | AKR1B7 | 7,450 ± 1,500 | 5.6 ± 0.35 | 0.001 | |
| NM_008012 | AKR1B8 | 635 ± 81 | 20 ± 0.52 | 0.031 | |
| NM_020299 | AKR1B10 | 7,800 ± 359 | 8.87 ± 0.63 | 0.002 | |
| NM_030611 | AKR1C3 | NDA | — | — | |
| NM_013778 | AKR1C6 | 12,073 ± 1,031 | 12.8 ± 0.67 | 0.001 | |
| NM_134066 | AKR1C18 | NDA | — | — | |
| NM_025337 | AKR7A5 | 5,931 ± 1,393 | 12.87 ± 0.27 | 0.002 | |
| Methylglyoxal | AKR1A4 | 874 ± 91 | 483 ± 0.27 | 0.55 | |
| AKR1B1 | 22 ± 2 | 37.2 ± 2.21 | 1.698 | ||
| AKR1B3 | 35 ± 4 | 19.5 ± 2.3 | 0.56 | ||
| AKR1B7 | 1,996 ± 274 | 8.9 ± 0.31 | 0.004 | ||
| AKR1B8 | 184 ± 15 | 18.2 ± 1.21 | 0.098 | ||
| AKR1B10 | 1,250 ± 89 | 26.82 ± 5 | 0.389 | ||
| AKR1C3 | NDA | — | — | ||
| AKR1C6 | 139 ± 35 | 7.65 ± 0.7 | 0.054 | ||
| AKR1C18 | NDA | — | — | ||
| AKR7A5 | 9,075 ± 1,250 | 22 ± 0.7 | 0.002 | ||
| Furfural | AKR1A4 | 5,918 ± 1,379 | 17.4 ± 0.78 | 0.002 | |
| AKR1B1 | 316 ± 34 | 30.86 ± 1.91 | 0.097 | ||
| AKR1B3 | 69 ± 10 | 13.36 ± 0.93 | 1.91 | ||
| AKR1B7 | NDA | — | — | ||
| AKR1B8 | 622 ± 100 | 2.6 ± 0.27 | 0.001 | ||
| AKR1B10 | 1,617 ± 287 | 178 ± 15 | 0.11 | ||
| AKR1C3 | 156 ± 35 | 3.09 ± 0.33 | 0.019 | ||
| Hydroxymethyl Furfural | AKR1A4 | 6,428 ± 1,479 | 24.05 ± 4 | 0.003 | |
| AKR1B1 | 482 ± 92 | 14.55 ± 2 | 0.03 | ||
| AKR1B3 | 98 ± 15 | 4.15 ± 0.3 | 0.042 | ||
| AKR1B7 | NDA | — | — | ||
| AKR1B8 | 1,197 ± 117 | 7.82 ± 0.05 | 0.016 | ||
| AKR1B10 | 1,117 ± 135 | 100 ± 15 | 0.085 | ||
| AKR1C3 | 1,939 ± 425 | 167 ± 0.02 | 0.008 | ||
| Deoxyglucosone | AKR1A4 | 2,067 ± 178 | 777 ± 25 | 0.376 | |
| AKR1B1 | 112 ± 5 | 38.9 ± 2 | 0.345 | ||
| AKR1B3 | 89 ± 5 | 11.6 ± 1 | 0.13 | ||
| AKR1B10 | 3,293 ± 118 | 14.36 ± 1.9 | 0.004 | ||
| AKR1B10 | 3,293 ± 118 | 14.36 ± 1.9 | 0.004 |
Enzyme activity was measured in 0.1 mol/l potassium phosphate (pH 7.0) using the indicated substrates and 0.15 mmol/l NADPH at room temperature. Proteins were reduced with dithiothreitol before assay. NDA, no detectable activity.