Table 1.

Computationally designed N- and C-terminal mutant CaMs

CaMs	Designed positions
	Site 1					Site 2					Site 3					Site 4
	20	22	24	27	31	56	58	60	62	67	93	95	97	100	104	129	131	133	135	140
CaM WT	D	D	D	I	E	D	D	N	T	E	D	D	N	I	E	D	D	D	Q	E
CaM-CWT	T	K	Q	I	E	S	D	E	I	K	–	–	–	–	–	–	–	–	–	–
CaM-NWT	–	–	–	–	–	–	–	–	–	–	A	R	D	M	E	R	D	N	E	N

All CaM residues that directly coordinate Ca²⁺ ions with their side chains (positions 20, 22, 24, 31, 56, 58, 60, and 67 for the N-terminal domain and 93, 95, 97, 104, 129, 131, 133, and 140 for the C-terminal domain) were included in the optimization. Residues 62 and 135 also were included because they coordinate Ca²⁺ ions with their backbone carbonyls. Residues 27 and 100, which do not coordinate Ca²⁺ ions, but are in close proximity to the ion and help define the Ca²⁺-binding loop, also were optimized. Bold lettering shows mutations with respect to the WT CaM sequence. –, positions were not included in the calculation for the mutant CaM indicated.