Table 2.

The energetic effect of phosphorylation and dephosphorylation of tyrosine residues (Y86, Y106, Y159, and Y187) in the TIR domain of TIRAP were calculated in terms of change in the free binding energy (BE) of TIRAP and p38 MAPK complexes.

Sr. No	TIRAP-p38 MAPK complex	Phosphorylated site(s) in TIRAP	Total BE (∆Gbind) (kcal/mol)	Change in ∆Gbind (kcal/mol)
A
1	TIRAP-p38 MAPK	–	− 10.0	–
2	p-Y86 and p38 MAPK	Y86	− 15.0	− 5
3	p-Y106 and p38 MAPK	Y106	− 10.9	− 0.9
4	p-Y159 and p38 MAPK	Y159	− 11.7	− 1.7
5	p-Y187 and p38 MAPK	Y187	− 12.0	− 2
6	p-all04 and p38 MAPK	Y86, Y106, Y159 and Y187	− 20.5	− 10.5

Sr. No	TIRAP-p38 MAPK complex	Dephosphorylated site in TIRAP	Total BE (∆Gbind) (kcal/mol)	Change in ∆Gbind (kcal/mol)
B
6	pYall04 and p38 MAPK	–	− 20.5	− 10.5
7	dpY86/pYall03 and p38 MAPK	Y86	− 7.1	− 13.4
8	dpY106/pYall03 and p38 MAPK	Y106	− 16.2	− 4.3
9	dpY159/pYall03 and p38 MAPK	Y159	− 9.6	− 10.9
10	dpY187/pYall03 and p38 MAPK	Y187	− 15.5	− 5

(A) Phosphorylation of tyrosine residues significantly decreases the binding energy (∆G_bind) of TIRAP and p38 MAPK complex, with the highest decrease seen with pY86 while the modest seen with pY106 (B) the sequential dephosphorylation shows the highest destabilizing effect occurs with dephosphorylation of Y86 and Y159 while the modest with Y106.