In silico engineering of aggregation-prone recombinant proteins for substrate recognition by the chaperonin GroEL

Table 2 Mutant Library Generated for ALLA_ECOLI and HCHA_ECOLI proteins of E.coli. The table shows a list of possible double mutants for wild type proteins and corresponding GRAVY values

S.No.	Swissprot ID's	Suggested Patches for Mutation (similar to GGIVLTG)	Patches after mutation	GRAVY value of mutated patch* (Compare with that of GroES mobile loop)**
1	ALLA_ECOLI	GDVIETQ	GI VII TQ	1.871
	ALLA_ECOLI	GDVIETQ	GI VIL TQ	1.771
	ALLA_ECOLI	GDVIETQ	GI VIV TQ	1.828
	ALLA_ECOLI	GDVIETQ	GL VII TQ	1.771
	ALLA_ECOLI	GDVIETQ	GL VIL TQ	1.671
	ALLA_ECOLI	GDVIETQ	GL VIV TQ	1.728
	ALLA_ECOLI	GDVIETQ	GV VII TQ	1.828
	ALLA_ECOLI	GDVIETQ	GV VIL TQ	1.728
	ALLA_ECOLI	GDVIETQ	GV VIV TQ	1.785
2	HCHA_ECOLI	GKLFSTG	GI LFI TG	2.014
	HCHA_ECOLI	GKLFSTG	GI LFL TG	1.914
	HCHA_ECOLI	GKLFSTG	GI LFV TG	1.971
	HCHA_ECOLI	GKLFSTG	GL LFI TG	1.914
	HCHA_ECOLI	GKLFSTG	GL LFL TG	1.814
	HCHA_ECOLI	GKLFSTG	GL LFV TG	1.871
	HCHA_ECOLI	GKLFSTG	GV LFI TG	1.971
	HCHA_ECOLI	GKLFSTG	GV LFL TG	1.871
	HCHA_ECOLI	GKLFSTG	GV LFV TG	1.928

As the single mutations for the patch doesn't make much difference in GRAVY value, so double mutations were considered.
*The GRAVY values are calculated by using ProtParam tool (ExPasy)
**The GRAVY value for mobile loop is = 1.514

ISSN: 1471-2164