Project name: DHFR [mutate: GR121A, CA85A, IA61A] [mutate: GA43A, GR96A, RA33A, MR20A, FA137A]

Chain sequence(s)	A: MMISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLDKPVIMGRHTWESSIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLPKKAQKKLYLTHIDDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCCFEILERR
Selected Chain(s)	A
Distance of aggregation	5 Å
FoldX usage	Yes
Dynamic mode	No
Automated mutations	No
Mutated residues	RA33A,MR20A,GR96A,FA137A,GA43A
Energy difference between WT (input) and mutated protein (by FoldX)	8.43788 kcal/mol CAUTION: Your mutation/s can destabilize the protein structure

Simulation log

[INFO]       Logger:   Verbosity set to: 2 - [INFO]                                                (00:00:01)
[WARNING]    runJob:   Working directory already exists (possibly overwriting previous results -ow 
                       to prevent this behavior)                                                   (00:00:01)
[INFO]       runJob:   Starting aggrescan3d job on: input.pdb with A chain(s) selected             (00:00:01)
[INFO]       runJob:   Creating pdb object from: input.pdb                                         (00:00:01)
[INFO]       FoldX:    Starting FoldX energy minimalization                                        (00:00:01)
[INFO]       FoldX:    Building mutant model                                                       (00:03:07)
[INFO]       FoldX:    Starting FoldX energy minimalization                                        (00:03:55)
[INFO]       Analysis: Starting Aggrescan3D on folded.pdb                                          (00:05:53)
[INFO]       Main:     Simulation completed successfully.                                          (00:05:55)
[INFO]       Logger:   Verbosity set to: 2 - [INFO]                                                (00:00:00)
[WARNING]    runJob:   Working directory already exists (possibly overwriting previous results -ow 
                       to prevent this behavior)                                                   (00:00:00)
[INFO]       runJob:   Starting aggrescan3d job on: input.pdb with A chain(s) selected             (00:00:00)
[INFO]       runJob:   Creating pdb object from: input.pdb                                         (00:00:00)
[INFO]       FoldX:    Starting FoldX energy minimalization                                        (00:00:00)
[INFO]       FoldX:    Building mutant model                                                       (00:02:43)
[INFO]       FoldX:    Starting FoldX energy minimalization                                        (00:03:44)
[INFO]       Analysis: Starting Aggrescan3D on folded.pdb                                          (00:04:55)
[INFO]       Main:     Simulation completed successfully.                                          (00:04:56)

• Minimal score value: -2.1876
• Maximal score value: 1.7419
• Average score: -0.4348
• Total score value: -69.1393

The table below lists A3D score for protein residues. Residues with A3D score > 0.0000 are marked by yellow rows.

residue index	residue name	chain	Aggrescan3D score	mutation
1	M	A	0.7532
2	I	A	0.3507
3	S	A	0.0000
4	L	A	0.0000
5	I	A	0.0000
6	A	A	0.0000
7	A	A	0.0000
8	L	A	0.0000
9	A	A	0.0000
10	V	A	0.2206
11	D	A	-1.7826
12	R	A	-0.8286
13	V	A	0.0000
14	I	A	0.0000
15	G	A	0.0000
16	M	A	-0.1637
17	E	A	-1.9245
18	N	A	-1.0853
19	A	A	-0.2896
20	R	A	-1.0770	mutated: MR20A
21	P	A	-0.2389
22	W	A	-0.1228
23	N	A	-1.2506
24	L	A	0.0000
25	P	A	-0.1333
26	A	A	-0.0110
27	D	A	0.0000
28	L	A	1.5882
29	A	A	0.3695
30	W	A	0.2503
31	F	A	0.2431
32	K	A	-1.2288
33	A	A	-0.2093	mutated: RA33A
34	N	A	-0.1715
35	T	A	0.0000
36	L	A	0.2568
37	D	A	-1.7479
38	K	A	-0.6681
39	P	A	0.0000
40	V	A	0.0000
41	I	A	0.0000
42	M	A	0.0000
43	A	A	-0.0788	mutated: GA43A
44	R	A	-0.7410
45	H	A	-1.0924
46	T	A	0.0000
47	W	A	0.0000
48	E	A	-1.8836
49	S	A	-0.2863
50	I	A	0.6038
51	G	A	-0.8633
52	R	A	-1.9361
53	P	A	-0.3306
54	L	A	0.1696
55	P	A	-0.2996
56	G	A	-0.5501
57	R	A	-0.5236
58	K	A	-1.2946
59	N	A	0.0000
60	I	A	0.0000
61	I	A	0.0000
62	L	A	0.1581
63	S	A	-0.0310
64	S	A	-0.4370
65	Q	A	-1.2847
66	P	A	-0.5355
67	G	A	-0.3732
68	T	A	-0.1901
69	D	A	-0.6770
70	D	A	-2.1876
71	R	A	-2.1707
72	V	A	0.0000
73	T	A	0.0147
74	W	A	0.1898
75	V	A	0.0000
76	K	A	-1.7167
77	S	A	-0.3484
78	V	A	-0.0830
79	D	A	-1.8780
80	E	A	-1.0573
81	A	A	0.0000
82	I	A	0.5635
83	A	A	0.0880
84	A	A	0.0236
85	C	A	0.0000
86	G	A	-0.7992
87	D	A	-1.7513
88	V	A	0.2940
89	P	A	-0.1657
90	E	A	-0.2291
91	I	A	0.0000
92	M	A	0.0000
93	V	A	0.0000
94	I	A	0.0000
95	G	A	0.0000
96	R	A	-0.2533	mutated: GR96A
97	G	A	0.0000
98	R	A	-1.8441
99	V	A	0.0000
100	Y	A	0.0000
101	E	A	-1.9079
102	Q	A	-0.8133
103	F	A	0.0000
104	L	A	0.1264
105	P	A	-0.4442
106	K	A	-1.2520
107	A	A	0.0000
108	Q	A	-0.8499
109	K	A	-0.4950
110	L	A	0.0000
111	Y	A	0.0000
112	L	A	0.0000
113	T	A	0.0000
114	H	A	-0.3420
115	I	A	0.0000
116	D	A	-1.7894
117	A	A	0.0000
118	E	A	-1.7750
119	V	A	-0.4394
120	E	A	-1.8118
121	G	A	-0.8547
122	D	A	-1.8298
123	T	A	-0.4548
124	H	A	-0.6094
125	F	A	0.0000
126	P	A	-0.3565
127	D	A	-1.7538
128	Y	A	-0.4340
129	E	A	-1.8262
130	P	A	-0.9037
131	D	A	-2.0196
132	D	A	-1.3288
133	W	A	0.0000
134	E	A	-1.2481
135	S	A	-0.0909
136	V	A	1.7419
137	A	A	0.3077	mutated: FA137A
138	S	A	-0.5766
139	E	A	-1.4746
140	F	A	1.5744
141	H	A	-0.2807
142	D	A	-1.8424
143	A	A	-0.6567
144	D	A	-1.7710
145	A	A	-0.4853
146	Q	A	-1.1877
147	N	A	0.0000
148	S	A	-0.2684
149	H	A	-0.3475
150	S	A	-0.3332
151	Y	A	0.0000
152	C	A	0.4090
153	F	A	0.0000
154	E	A	-0.2244
155	I	A	0.2313
156	L	A	0.0000
157	E	A	-0.8310
158	R	A	-0.9913
159	R	A	-1.9401