TABLE 2.

Comparison of TSC1-TBC1D7 binding (pulldown versus co-IP): TSC1 and TBC1D7 amino acid substitutions

	Effect on TSC1-TBC1D7 interaction
	Pulldown	TBC1D7 IP; TSC1 co-IP (Fig. 6g)	TSC1 IP; TBC1D7 co-IP (Fig. 6e)
TSC1 substitution
I954A (helix A)	No interaction	Reduced interaction	Reduced interaction
F958A (helix A)	No interaction	Reduced interaction	Reduced interaction
I962A (helix B)	No interaction	Reduced interaction	Reduced interaction
L965A/L969A (helix A/B)a	Reduced interaction	Reduced interaction	No significant reduction
Y966A (helix B)	Reduced interaction	Reduced interaction	No significant reduction
TBC1D7 substitution
R81A/Q84A/R121A	No interaction	No interaction	Reduced interaction
H90A/R96A	No interaction	No significant reduction	Reduced interaction
V94A/V95A	No interaction	Reduced interaction	Reduced interaction
R96A	Reduced interaction	No significant reduction	No significant reduction
L114A	No interaction	No significant reduction	No significant reduction
R121A	No significant reduction	No significant reduction	No significant reduction

^a The two L965/L969 pairs from both TSC1 helices interact with each other and play a role in stabilizing TSC1 dimerization, but neither residue directly contacts TBC1D7.