Table 1.
HiSol scores and conserved residues corresponding to mutated residues in MeHNL, SuPDH, ChMOX, AtADC, and MpLUC.
| Enzyme | Target residue | How residues were identified | HiSol score (Ranking)a | Conserved residuesb | Positionc | Result |
|---|---|---|---|---|---|---|
| SuPDH | Lys148 | Hydropathy contradiction rule d | −2.748 (1st score) | Ile (64%), Val (21%), Leu (9%) | Coil | Negative |
| Gln225 | −2.232 (2nd score) | Val (73%) | Helix | Positive | ||
| Lys374 | −2.023 (3rd score) | Ile (53%) | Coil | Negative | ||
| Gln337 | −1.949 (4th score) | Ile (72%) | Helix | Positive | ||
| Gln243 | −1.807 (5th score) | Val (62%) | Coil | Negative | ||
| AtADC | Leu435 | α-helix rulee | 1.814 (8th score) | His (31%) | Helix | Positive |
| Lys441 | −1.334 (11th score) | Leu (21%), Lys (11%) | Helix | False negative | ||
| MeHNL | His103 | Hydropathy contradiction ruled | −2.283 (1st score) | Leu (47%), Val (30%) | Sheet | False negative |
| ChMOX | Val444 | α-helix rulee,f | 1.834 (10th score) | Glu (9%) | Helix | False negative |
| Val455 | 0.621 (106th score) | Leu (42%), Ile (18%) | Helix | False negative | ||
| DmODC | Lys117 | α-helix rulee | −1.795(3rd score) | Leu (30.5%), Ala (22.5%), Tyr (13.3%) | Helix | Positive |
| DmGDH | Val174 | α-helix rulee | 2.058 (3rd score) | Asp (24.3%), Glu (15.8%), | Helix | Positive |
| MpLUC | Ile80 | Hydropathy contradiction rule on α-helixg | 1.919 (2nd score) | Lys (53%), Ile (17%) | Helix | Positive |
| Arg87 | −1.293 (10nd score) | Arg (41%), Lys (20%), Val (11%), Phe (11%) | Helix | Negative | ||
| Ala177 | 1.361 (7th score) | Asp (57%), Ala (20%), Glu (13%) | Helix | Positive | ||
| hGH | Leu114 | Hydropathy contradiction rule on α-helixg | 2.042 (1st score) | Lys (41.4%) | Helix | Positive |
| Arg95 | −1.473 (2nd score) | Ser (41.1%), Val (16.9%) | Helix | Positive | ||
| Leu46 | 1.463 (3rd score) | Lys (12.2%), Phe (10.7%) | Helix | Positive | ||
| Leu88 | 1.369 (4th score) | Leu (33.2%), Glu (19.2%), Asn (16.0%) | Helix | Negative | ||
| Phe55 | 1.319 (5th score) | His (30.6%), Phe (20.2%), Asn (14.9%) | Helix | Positive | ||
| Leu82 | 1.231 (6th score) | Leu (23.8%), Arg(19.8%), Val (10.0%) | Helix | Negative | ||
| Val97 | 1.191 (7th score) | Val (28.3%), Glu (16.2%) | Helix | Positive |
aRanking of HiSol score is represented for the predicted hotspots. Here, the score bearing negative value would be ranked after lining the scores with descending order, and vice versa for the score bearing positive value.
bRatio of conservation > 10%.
cPosition in secondary structure was predicted by PSIPRED.
d“Hydropathy contradiction rule” means that the hotspots were predicted based on the analysis of the HiSol scores.e“α-helix rule” means that the hotspots were predicted from the α-helix rule according to the hydropathy index of the residues. The HiSol scores were not used for the prediction, but are shown in italic as references. Thus, the ranking of the HiSol scores are not always high, but distributed sparse (from 3rd to even 106th).fThese are good examples showing the merit of theα-helix rule: even residues with low appearance rates and lower HiSol scores were chosen as hotspots (V444E with 10th HiSol score and only 9% appearance rate, and Val455 with 106th HiSol score in ChMOX). g“Hydropathy contradiction rule on α-helix” means that hotspot residues were first selected based on HiSol scores, and then the residues located on the α-helices were selected.