Papers by Sara Bashraheel
International Journal of Molecular Sciences
Bacterial superantigens (SAgs) are effective T-cell stimulatory molecules that lead to massive cy... more Bacterial superantigens (SAgs) are effective T-cell stimulatory molecules that lead to massive cytokine production. Superantigens crosslink between MHC class II molecules on the Antigen Presenting Cells (APC) and TCR on T-cells. This enables them to activate up to 20% of resting T cells, whilst conventional antigen presentation results in the activation of 0.001–0.0001% of the T cell population. These biological properties of superantigens make them attractive for use in immunotherapy. Previous studies have established the effectiveness of superantigens as therapeutic agents. This, however, was achieved with severe side effects due to the high lethality of the native toxins. Our study aims to produce superantigen-based peptides with minimum or no lethality for safer cancer treatment. In previous work, we designed and synthesized twenty overlapping SPEA-based peptides and successfully mapped regions in SPEA superantigen, causing a vasodilatory response. We screened 20 overlapping SPE...
Biomedicine & Pharmacotherapy
<p>A) Combined CD spectra data of Xen CPG2 and Ps CPG2 in molar ellipticity relative the wa... more <p>A) Combined CD spectra data of Xen CPG2 and Ps CPG2 in molar ellipticity relative the wavelength in far UV region, the spectra obtained by dragging their spectral data over each other, where smooth 0 is molar ellipticity of Xen CPG2 and smooth 1 is for CD spectra of Ps CPG2. All Spectral data are corrected for the baseline buffer, B) represents the combined High voltage (HV) for both enzymes where average 0 is Xen CPG2 and average 1 is Ps CPG2. Also the table shows the calculated protein secondary structure of Xen CPG2 and Ps CPG2 by CDNN deconvolution analysis using their CD spectral data.</p
<p><b>A.</b> Electrostatic surface presentation of the Xen CPG2 tetramer. Posit... more <p><b>A.</b> Electrostatic surface presentation of the Xen CPG2 tetramer. Positive and negative charges are shown in blue and red, respectively. <b>B.</b> Color rendering of a Xen CPG2 monomer. Helix in cyan, b-sheet in pink, loops in brown and amino acids which differ from the model carboxypeptidase G2 are shown as yellow sticks. Rendering was performed using PyMol. <b>C.</b> Stereoview of the alignment of CPG2 Pseudomonas sp. Strain RS-16 (blue cartoon, PDB ID 1CG2) with the model of Xen CPG2 (green cartoon, RMS = 0.084 (374 to 374 atoms)).</p
<p><b>a</b>. Coomassie blue staining of 10% SDS-PAGE of <i>E</i>. &... more <p><b>a</b>. Coomassie blue staining of 10% SDS-PAGE of <i>E</i>. <i>coli</i> BL21(DE3)RIL-CPG2 protein expression at 37°C of the gene isolated from <i>Xenophilus azovorans</i> SN213. M is the PageRuler Prestained Protein Ladder (10 to 180 kDa), lanes 1 and 2 are the induced soluble and insoluble fractions respectively, and lanes 3 and 4 are the uninduced soluble and insoluble fractions respectively. <b>b</b>. Protein expression at 20°C of the gene isolated from <i>Xenophilus azovorans</i>. SN213. M is the prestained protein ladder, lanes 1 and 2 are the induced soluble and insoluble fractions respectively, and lanes 3 and 4 are the uninduced soluble and insoluble fractions respectively.</p
<p>a) DAMPA H+ peak is the product of folate hydrolysis by the isolated strain. b and c) P1... more <p>a) DAMPA H+ peak is the product of folate hydrolysis by the isolated strain. b and c) P1, P2 are DMPA H+ produced by recombinant CPG2s, new and Ps CPG2 respectively and F1, F2 are intact folate isolated from the media of both recombinant enzymes.</p
<p><b>A.</b> Dot blot using anti His tag antibody and using anti Xen CPG2 antib... more <p><b>A.</b> Dot blot using anti His tag antibody and using anti Xen CPG2 antibody where 1, 2, and 3 are pure protein (Xen CPG2 and Ps CPG2) at concentrations (0.05, 0.1, and 0.2 mg/mL). <b>B.</b> Dot blot at different concentration of anti Xen CPG2 antibody where 1, 2, and 3 are blotting at dilutions 1:20 000, 1:10 000 and 1:3000 in blocking buffer. <b>C.</b> SDS-PAGE and Western blot analysis of the pure protein (Xen CPG2 and Ps CPG2) where M is PageRuler™ Unstained Protein Ladder (10–200 kDa), lanes 1, 2, 3 are 0.25, 0.1, and 0.05 mg/mL of Xen CPG2 and lanes 4, 5, 6 are the same series of protein concentrations of Ps CPG2.</p
<p>MTX substrate solution and pure CPG2 in the presence of Zn<sup>2+</sup> (sta... more <p>MTX substrate solution and pure CPG2 in the presence of Zn<sup>2+</sup> (star), control reaction of MTX substrate solution and buffer (filled triangle) and MTX substrate solution and pure CPG2 in the presence of Zn<sup>2+</sup> and 10 mM EDTA (square) are shown. New isolated CPG2 is Zn<sup>2+</sup> dependent.</p
<p>Coomassie blue staining of a 10% SDS-PAGE gel. M; Size markers in kiloDaltons; M1 is Pag... more <p>Coomassie blue staining of a 10% SDS-PAGE gel. M; Size markers in kiloDaltons; M1 is PageRuler Prestained Protein Ladder (10 to 180 kDa) and M2 is SeeBlue Plus prestained standard (↱3 to 198 kDa). <b>a)</b> Xen CPG2 purification; lane 1 is total soluble fraction of glucarpidase after centrifugation; 2, flow through; 3–4, wash 1 and wash 5; 5–9, eluted fractions from the Ni-NTA column with 400 mM imidazole. <b>b)</b> Ps CPG2 purification; lanes 1, 2, 3 are total, flow through, wash, lanes 4–6 are elution fractions.</p
<p>MTX substrate solution and total soluble protein of <i>Pseudomonas lubricants</... more <p>MTX substrate solution and total soluble protein of <i>Pseudomonas lubricants</i> strain SF168 in presence of Zn<sup>2+</sup> (filled ball) and in presence of Zn<sup>2+</sup> and EDTA (diamond). MTX substrate solution and total soluble protein from <i>Xenophilus</i> sp. SN213 in the presence of Zn<sup>2+</sup> (star), and in the presence of Zn<sup>2+</sup> and EDTA (filled triangle), and control of MTX and enzyme buffer (square) are shown. The data in this figure indicates that both strains (<i>Pseudomonas lubricans</i> strain SF168 and <i>Xenophilus</i> sp. SN213) show CPG2 activity through methotrexate hydrolysis.</p
<p>The activity of the isolated recombinant CPG2 in comparison to Ps CPG2 of <i>Pseud... more <p>The activity of the isolated recombinant CPG2 in comparison to Ps CPG2 of <i>Pseudomonas sp</i> strain RS-16 and in the presence of negative control (vector only) on LB/KAN/CAM/IPTG/Folate in the presence of 0.2 mM ZnSO<sub>4</sub>.</p
Biomedicine & Pharmacotherapy, 2020
Biomedicine & Pharmacotherapy, 2019
Biomedicine & Pharmacotherapy, 2019
European Journal of Pharmaceutical Sciences, 2019
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Papers by Sara Bashraheel