Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.

<p>Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.</p

Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.

<p>Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.</p

List of human plasma proteins with mass spectrometric molar intensities similar to those from the most abundant trypanosome proteins found in plasma from sleeping sickness patients.

*<p>Protein concentrations were retrieved from reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Hortin1" target="_blank">[19]</a>. Human proteins reported to be depleted by the LC20 IgY14 and Supermix LC10 columns <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Patel1" target="_blank">[26]</a> have been omitted from this table.</p

Summary of patient information.

<p>Plasma and CSF were collected from <i>T. b. rhodesiense</i>-infected patients confirmed to have late stage human African trypanosomiasis at Lwala Hospital in central Uganda.</p

ELISA detection of trypanosome antigens in plasma from <i>T. b. rhodesiense</i> -infected patients with late-stage human African trypanosomisis.

<p>All samples were tested in duplicate and the average response is shown. Error bars represent 1 standard deviation.</p

Proteases and ubiquitin proteins discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.

<p>Proteases and ubiquitin proteins discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.</p

Primary sequence alignment of <i>T</i>. <i>congolense</i> calflagin with those of <i>T</i>. <i>cruzi</i> FCaBP and <i>T</i>. <i>brucei</i> Tb24.

<p>The secondary structural elements (α-helices and β-strands) are depicted as cones and arrows, respectively, and were derived from the I-TASSER based structure prediction for <i>T</i>. <i>congolense</i> calflagin, the x-ray crystal structure of <i>T</i>. <i>cruzi</i> FCaBP (3CS1) and the NMR data for <i>T</i>. <i>brucei</i> Tb24 (2LVV). The four EF-hands (EF1, EF2, EF3, and EF4) are highlighted in green, salmon, cyan, and yellow, respectively. Residues in the 12-residue Ca²⁺ binding loops at position 1, 3, 5 and 12 are underlined. Invariant basic residues on the protein surface that are associated with membrane binding are colored blue. Non-conserved surface-exposed residues are highlighted using bold print.</p

Multiple sequence alignment of the <i>T</i>. <i>congolense</i> calflagins.

<p>The positions of the two MS-identified tryptic peptides identified by mass spectrometry are highlighted in yellow and red boxes. The lower case, white highlighted v represent amino acid (valine-isoleucine) differences in two of the ORFs.</p

Calflagin-based serodiagnosis of trypanosome infections in Ugandan cattle.

<p>(A) ELISA signal intensities resulting from bovine IgG and IgM antibodies binding to solid-phase adsorbed recombiant <i>T</i>. <i>congolense</i> calflagin. All values were normalized against signals elicited from plasma of laboratory raised, trypanosome negative calves. (B) ROC curves generated from the serodiagnostic ELISA results. Areas under curve for IgG and IgM were 0.623 and 0.709 respectively. For reference, an area under the curve of 1.00 equates to a test with 100% sensitivity and specificity, whereas an area under the curve of 0.500 indicates that a test has no value in differentiating between the binary population.</p

Structural characterization and surface analysis of <i>T</i>. <i>congolense</i> calflagin Modeled <i>T</i>. <i>congolense</i> calflagin (middle panel) was compared to the crystal structure of <i>T</i>. <i>cruzi</i> FCaBP (left vertical column) and to the NMR structure of <i>T</i>. <i>brucei</i> Tb24 (right vertical column).

<p>(A) the predicted model of <i>T</i>. <i>congolense</i> calflagin (middle vertical column) exhibits four EF-hands motifs. The EF-hands (EF1, EF2, EF3 and EF4) are colored green, salmon, cyan, yellow, accordingly. Left panel: structural alignment of <i>T</i>. <i>congolense</i> (grey) over <i>T</i>. <i>cruzi</i> FCaBP (green). Right panel: <i>T</i>. <i>congolense</i> calflagin structure aligned over <i>T</i>. <i>brucei</i> Tb24 (magenta). (B) Surface representation of the three calflagin models overlapping panel A, showing exposed hydrophobic (gray), basic (blue) and acidic (red) respectively. (C) 180° rotation of B in the <i>y-axis</i>. Black arrows pointing towards putative epitopes and numbered according to their respective α-helix location.</p