. Author manuscript; available in PMC: 2017 Aug 30.

Published in final edited form as: J Mol Biol. 2016 Feb 23;428(8):1515–1530. doi: 10.1016/j.jmb.2016.02.020

Table 1.

Characterization of ETV6 variants

	Helix H5 sequence^a	AGADIR^b (%)	L337 PF^c	H5 Avg. PF^d	ΔΔG_HX (kcal/mol)^e	K_D (nM)^f	Fold-increase inhibition^g
ETV6^ΔH5	No helix H5		nd^h			0.1ⁱ
ETV6^WT	⁴²⁷TDRLEHLESQVLDE⁴⁴⁰	0.4	nd^h	13		6 ± 1	1
ETV6^E434Q	TDRLEHLQSQVLDE	0.6	nd^h		Unfolded H5
ETV6^E434D	TDRLEHLDSQVLDE	0.4	nd^h		Unfolded H5
ETV6^H432R	TDRLERLESQVLDE	1.6	11	17	0.1	30 ± 10	5
ETV6^RAE	TDRLERLEAEVLDE	3.2	20	23	0.3	90 ± 10	15
ETV6^2C-ox	TDRLEHLESQCLDE	0.4	116	68	1.0	20,000 ± 9000^j	3000

The amino acid substitutions are bold underlined.

AGADIR predicted helical propensity for each given sequence as an isolated polypeptide [17].

Measured at 35 °C and 11.7 T (500 MHz) NMR field strength.

Measured at 30 °C and pH values of 6.4 and 7.2. PFs of residues 430 to 440 were averaged.

$Δ Δ G_{HX} = Δ G_{HX}^{mut} - Δ G_{HX}^{Wt}$ , where ΔG_HX = RTln(PF) for unfolding to an exchange competent state was calculated from the average PFs for the helix H5 residues 430 to 440.

Measured by EMSA with a 23-bp DNA duplex containing the ETS recognition sequence ^5′−GGAA^−3′. The data are the average ± standard deviation of at least four measurements.

Fold increase in autoinhibition with respect to inhibited ETV6^WT.

Not detected.

ⁱ

Value reported [13].

Measured by NMR-based titrations using a 15-bp DNA containing a consensus ^5′−GGAA^−3′ sequence.