Additional file 3: of Individual odour signatures that mice learn are shaped by involatile major urinary proteins (MUPs)

GC-MS analysis used for Fig. 6. Peak areas and principal component scores for the four male-specific volatiles measured from BALB/c urine, C57BL/6 urine and BALB/c urine + r-darcin. (XLS 31 kb

Additional file 1: of Individual odour signatures that mice learn are shaped by involatile major urinary proteins (MUPs)

Intact mass spectra and GC-MS data from trios of wild male brothers used for Figs. 4 and 5. Figure 4AB tab provides intact mass spectra (expressed as proportion of highest peak) for each male according to sib group and genotype. Figure 4CD tab provides areas for 134 volatiles peaks (expressed as % total peak area) and principal component analysis. (XLS 232 kb

Additional file 2: of Individual odour signatures that mice learn are shaped by involatile major urinary proteins (MUPs)

Datasets from behavioural recognition tests in Figs. 1, 2 and 3. Time spent under male and female stimuli for each test and replicate together with summary data and statistical comparison. (XLS 67 kb

Comparison of binding cavities of darcin and MUP11.

<p>Left: Overlay of binding residues of darcin (mauve) and MUP11 (orange), where the differing amino acid residues in both darcin/MUP11 are labelled. Right: schematic of residues highlighted as part of the SBT binding site, conserved residues between darcin and MUP11 are green and variable residues are coloured red with the darcin residue only indicated. Bottom: Aligned sequences with SBT binding residues highlighted using the same colour scheme as above (conserved  =  green; variable  =  red), secondary structure schematic is aligned below the sequences with identical colour scheme to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108415#pone-0108415-g002" target="_blank">Fig. 2</a>.</p

MUP cavity analysis.

<p>A binding cavity consensus was determined based on the active ligands identified by LigPLOT <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108415#pone.0108415-Wallace1" target="_blank">[49]</a> and PDBePISA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108415#pone.0108415-Krissinel1" target="_blank">[48]</a> in over 50% of the complex structures and shown mapped (in yellow) onto darcin (2L9C) and MUP11 (2LB6) (top two sequences). All ligand:MUP complexes available in the PDB are analysed. Residues identified as part of the binding site are highlighted according to ligand type: aromatic/pyrazole ligand (blue), aliphatic/non-cyclic molecule (orange).</p

Variation in surface amino acids between darcin and MUP11.

<p>Darcin (mauve) (A) and MUP11 (orange) (B) are shown in the same orientations. Non-conserved surface exposed residue side-chains are shown as stick representations and shaded cyan (darcin) and red (MUP11). Only variations of residues that do not confer similar properties (polar, hydrophobic, charged, aromatic etc.) are shown, as Patches 1, 2 and 3 (see text). For clarity hydrogen atoms are omitted from the stick-representations of the residues shown.</p

Isothermal titration calorimetry curves.

<p>Plots showing 2-<i>sec</i>-butyl thiazole (SBT) binding to darcin and MUP11 in 25 mM PO₄³⁻, 25 mM NaCl, 298 K curve fit to a one-site model. (A) Darcin binds SBT with N (stoichiometry ratio)  =  1.0, K_D∼0.173 µM, ΔH = <b>∼</b> -13.1 kcal/mol and TΔS = ∼3.9 kcal/mol. (B) MUP11 binds SBT with N (stoichiometry ratio)  = 1.0, K_D ∼2.76 µM, ΔH ∼-9.8 kcal/mol and TΔS = ∼ 2.2 kcal/mol.</p

Structural statistics for the refined NMR structures of darcin and MUP11.

<p>*Quoted Root-Mean-Square Deviation (RMSD) is derived from comparison of closest-to-mean structure; i.e. representative structure, of each ensemble.</p><p>Structural statistics for the refined NMR structures of darcin and MUP11.</p

Structural features of Darcin and MUP11^(a).

(a)<p>As defined by the programme Stride <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108415#pone.0108415-Heinig1" target="_blank">[40]</a>.</p>(b)<p>Loop nomenclature: L₁ (between β1 and β2), L₂ (β2-β3), L₃ (β3-β4), L₄ (β4-β5), L₅ (β5-β6), L₆ (β6-β7), L₇ (β7-β8), L₈ (β8-β9), L₉ (α1-β9).</p>(c)<p>Conserved C-terminal 3₁₀-helix.</p><p>Structural features of Darcin and MUP11^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108415#nt102" target="_blank">(a)</a>}.</p

Schematic of MUP beta-barrel and inter-strand loop arrangement.

<p>Top left: top-down view of the beta barrel. Loops at the top, N-terminal end of the barre<b>l</b> are highlighted in green and magenta. Top right: bottom-up (C-terminal end,) view of the beta barrel. Loops at the bottom, C terminal end of the barrel are highlighted in blue and tan. Bottom: Alignment of darcin and MUP11 sequences with paired loop residues used to measure inter-loop distances highlighted, each residue pairs are coloured green, magenta, tan and blue in accordance with the schematic views.</p