Table 3.

Site-directed mutants of adrenocorticotropic hormone receptor (ACTHR) and their effects on receptor function.

Mutation	Binding (×wtACTHR)	Cyclic adenosine 3′,5′-monophosphate (×wtACTHR)	Membrane trafficking (×wtACTHR)	Rate of internalization (% of initial rate)	Reference
D70 → A	X	X	X		(146)
*K77 → A	Ki = 4.5×↑	EC50 = 7.8×↑	1.4×↓
E80 → A	Ki = 11.6×↑	EC50 = 9.8×↑	1.3×↓
*N81 → A	Ki = 1.8×↑	EC50 = 2.4×↑	1.1×↓
D103 → N	Ki = 2.8×↑	EC50 = 7.3×↑	1.5×↓
*D104 → N	Ki = 10.2×↑	EC50 = 18.3×↑	1.3×↓
D107 → N	Ki > 178.6×↑	EC50 > 1250.0×↑	1.5×↓
*F110 → A	Ki > 116.3×↑	EC50 > 208.3×↑	1.5×↓
*T164 → A	Ki = 1.2×↓	EC50 = 1.3×↓	1.1×↓
*F168 → A	Ki = 25.3×↑	EC50 = 68.3×↑	1.8×↓
*F178 → A	Ki > 178.6×↑	EC50 > 1250.0×↑	1.5×↓
F235 → A	Ki = 17.5×↑	EC50 = 270.0×↑	1.2×↓
H238 → A	Ki > 178.6×↑	EC50 > 1250.0×↑	1.4×↓
*F244 → A	Ki = 1×↔	EC50 = 1.1×↑	1.1×↓
F258 → A	Ki = 13.9×↑	EC50 = 233.8×↑	1.3×↓
D272 → A	X	X	X
N12N13 → QQ		EC50 = ~1.4×↑	↔		(194)
N17N18 → QQ		EC50 = ~2.1×↑	↔
N12N13N17N18 → QQQQ		EC50 = ~4.5×↑	~3.2×↓
wtACTHR		Rmax = 1×↔	1×↔	33.1	(86)
T131 → A		Rmax = 1.1×↓	1.27×↑	<0.1
T131 → D		Rmax = 4.5×↓	1.3×↑	<0.1
S140 → A		Rmax = 1.1×↓	1.1×↓	25.5
S140 → D		Rmax = 1.2×↓	1.25×↑	43.2
T143 → A		Rmax = 33.3×↓	100×↓	22.2
T143 → D		Rmax = 1×↔	3.1×↓	16.3
T143 → S		Rmax = 1.1×↑	2.3×↓	27.7
T143 → G		X	11.1x↓	X
T143 → K		Rmax = 2.9×↓	12.5×↓	X
T147 → A		Rmax = 1.2×↑	1.2×↑	23.6
T147 → D		Rmax = > 100×↓	>100×↓	<0.1
S202 → A		Rmax = 1×↔	1.5×↑	40.6
S202 → D		Rmax = 1.1×↑	1.1×↓	60.7
S204 → A		Rmax = 1.2×↑	1×↔	18.8
S204 → D		Rmax = 1×↔	1.5×↓	39.4
S208 → A		Rmax = 1.1×↑	1.1×↓	22.6
S208 → D		Rmax = 1.1×↓	1.1×↓	56.7
T209 → A		Rmax = 1.1×↑	1×↔	32.4
T209 → D		Rmax = 1.1×↓	1.2×↓	20.1
S280 → A		Rmax = 1×↔	1.4×↓	50.8
S280 → D		Rmax = 1.5×↓	1.1×↓	<0.1
S294 → A		Rmax = 1×↔	1.3×↓	36.4
S294 → D		Rmax = 1.4×↓	1.3×↓	49.0
N12N13N17N18 → QQQQ		Rmax = 1.2×↓	3.0×↓	80.5

×, fold difference compared with wtACTHR; ↔, value unaffected; ↑, value increased; ↓, value decreased; X, not determined; empty field, no data available.