3 research outputs found
400 μM OdDHL treatment significantly increases necrosis of MDA-MB-231 cells in all conditions.
<p>(A) Differences in the annexin V-FITC staining mean gray value (MGV) for MDA-MB-231 cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3). (B) Differences in the propidium iodide staining MGV for MDA-MB-231 cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01, * = p-value < 0.05 based on least mean contrasts. (C) Differences in the annexin V-FITC staining MGV for MCF-DCIS.com cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia (N = 3), 2D/hypoxia (N = 4), and 2D/normoxia culture conditions (N = 4); *** = p-value < 0.001 based on least mean contrasts. (D) Differences in the propidium iodide staining MGV for MCF-DCIS.com cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01, * = p-value < 0.05 based on least mean contrasts. (E) Differences in the annexin V-FITC staining MGV for MCF-10A cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01 based on least mean contrasts. (F) Differences in the propidium iodide staining MGV for MCF-10A cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01 based on least mean contrasts. Error bars represent standard error of the mean.</p
Chemical Proteomic Platform To Identify Citrullinated Proteins
Anti-citrullinated protein antibodies
(ACPAs) are a hallmark of
rheumatoid arthritis (RA) and are routinely used for disease diagnosis.
Protein citrullination is also increased in cancer and other autoimmune
disorders, suggesting that citrullinated proteins may serve as biomarkers
for diseases beyond RA. To identify these citrullinated proteins,
we developed biotin-conjugated phenylglyoxal (biotin-PG). Using this
probe and our platform technology, we identified >50 intracellular
citrullinated proteins. More than 20 of these are involved in RNA
splicing, suggesting, for the first time, that citrullination modulates
RNA biology. Overall, this chemical proteomic platform will play a
key role in furthering our understanding of protein citrullination
in rheumatoid arthritis and potentially a wider spectrum of inflammatory
diseases
Biological Studies and Target Engagement of the 2‑<i>C</i>‑Methyl‑d‑Erythritol 4‑Phosphate Cytidylyltransferase (IspD)-Targeting Antimalarial Agent (1<i>R</i>,3<i>S</i>)‑MMV008138 and Analogs
Malaria
continues to be one of the deadliest diseases worldwide, and the emergence
of drug resistance parasites is a constant threat. <i>Plasmodium</i> parasites utilize the methylerythritol phosphate (MEP) pathway to
synthesize isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate
(DMAPP), which are essential for parasite growth. Previously, we and
others identified that the Malaria Box compound MMV008138 targets
the apicoplast and that parasite growth inhibition by this compound
can be reversed by supplementation of IPP. Further work has revealed
that MMV008138 targets the enzyme 2-<i>C</i>-methyl-d-erythritol 4-phosphate cytidylyltransferase (IspD) in the
MEP pathway, which converts MEP and cytidine triphosphate (CTP) to
cytidinediphosphate methylerythritol (CDP-ME) and pyrophosphate. In
this work, we sought to gain insight into the structure–activity
relationships by probing the ability of MMV008138 analogs to inhibit <i>Pf</i>IspD recombinant enzyme. Here, we report <i>Pf</i>IspD inhibition data for fosmidomycin (FOS) and 19 previously disclosed
analogs and report parasite growth and <i>Pf</i>IspD inhibition
data for 27 new analogs of MMV008138. In addition, we show that MMV008138
does not target the recently characterized human IspD, reinforcing
MMV008138 as a prototype of a new class of species-selective IspD-targeting
antimalarial agents