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Pairwise interaction scores highlight two activation pathways. There are two critical stabilizing routes between the α1 and α5 helices in the GDP-bound state. The first (purple) is between Gln52 (α1), Ile56 (α1), and Thr329 (α5). The second pathway (blue) connects Met53 (α1), His57 (α1), Val332 (α5), and Phe336 (α5). A, WT Gαi1 (PDB entry 1GIA (26)) maintains both networks in the GDP-bound state. B, the Ins4A mutant loses the hydrophobic core between α5, α1, and the β2-β3 strands.
