Characterisation of the interaction between NOXO1 and NOXA1.

<p>(<b>A</b>) Upper part shows the raw calorimetric data for the interaction between human NOXA1 SH3 and NOXO1 SH3_AB–E. Lower part shows the integrated heat changes, corrected for heat of dilution, and fitted to a single site binding model. (▪) Human NOXA1 SH3 titrated into human NOXO1 SH3_AB–E (▴) Mouse NOXA1 SH3 titrated into mouse NOXO1 SH3_AB–E. (<b>B</b>) Upper part shows raw calorimetric data for the interaction of peptideA and human NOXA1 SH3. Lower part shows the integrated heat changes, corrected for heat of dilution, and fitted to a single site binding model. (▪) PeptideA titrated into human NOXA1 SH3 (▴) PeptideA titrated into mouse NOXA1 SH3.</p

Characterisation of the interaction between NOXO1 SH3_AB–E and p22^phox.

<p>Upper part shows the raw calorimetric data for the interaction of mouse p22^phoxC- NOXO1 SH3_AB–E. Lower part shows the integrated heat changes, corrected for heat of dilution, and fitted to a single site binding model. (▪) Human p22^phoxC titrated into human NOXO1 SH3_AB–E (▴) Mouse p22^phoxC titrated into mouse NOXO1 SH3_AB–E.</p

Characterisation of the interaction between the tandem SH3 domains of NOXO1 and peptides p22^phox.

<p>Upper part shows the raw calorimetric data for the interaction of p22^phoxC and human NOXO1 SH3_AB. Lower part shows the integrated heat changes, corrected for heat of dilution, and fitted to a single site binding model. (▪) Human p22^phoxC titrated into human NOXO1 SH3_AB (▴) Mouse p22^phoxC titrated into mouse NOXO1 SH3_AB.</p

Characterisation of intermolecular interactions between NOXO1 and NOXA1.

<p>All measurements were performed at 18°C. The <i>K</i>_d is given in units of 10⁻⁶ M, Δ<i>H</i> and TΔ<i>S</i> are given in kcal mol⁻¹ (1 kcal/mol≡4.184 kJ/mol). The stoichiometry of complex formation for each binding site is N = 1.0±0.1.</p

Characterisation of intermolecular interactions between NOXO1 and p22^phox.

<p>All measurements were performed at 18°C. The <i>K</i>_d is given in units of 10⁻⁶ M, Δ<i>H</i> and TΔ<i>S</i> are given in kcal mol⁻¹ (1 kcal/mol≡4.184 kJ/mol). The stoichiometry of complex formation for each binding site is N = 1.0±0.1. No binding is indicated by NB.</p

Schematic representation of the domain structures of p47^phox, NOXO1, p67^phox and NOXA1.

<p>(<b>A</b>) Predicted domain structures of NOXO1 and NOXA1 in comparison to p47^phox and p67^phox, respectively. Human and mouse constructs used in this study are illustrated by black lines. Mouse constructs are identical in length unless otherwise stated in brackets. The autoinhibitory region (AIR) and proline rich region (PPR) are indicated. (<b>B</b>) Alignment of p47^phox, human NOXO1 and mouse NOXO1. The PX domains (grey shaded), SH3 domains (cyan shaded), polybasic region (orange shaded ox) and proline rich motif (green shaded) are indicated. (<b>C</b>) Structure of the autoinhibited core of p47^phox showing the superSH3 domain conformation. The structure shows the biologically relevant monomeric form of the protein that is also observed in solution, not the domain-swapped crystallized dimer <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0010478#pone.0010478-Groemping2" target="_blank">[22]</a>. The SH3 domains and polybasic region are highlighted in cyan and orange, respectively.</p