A5061293261- Bone health and treatments
- Chronic Myeloid Leukemia Treatments
- Endoplasmic Reticulum Stress and Disease
- Ubiquitin and proteasome pathways
- Mitochondrial Function and Pathology
- Synthesis and biological activity
- Retinoids in leukemia and cellular processes
- Acute Myeloid Leukemia Research
- Protein Kinase Regulation and GTPase Signaling
- Cancer-related Molecular Pathways
- Protein Degradation and Inhibitors
- Cancer, Hypoxia, and Metabolism
- Autophagy in Disease and Therapy
- Heat shock proteins research
- MicroRNA in disease regulation
- Circular RNAs in diseases
- Transplantation: Methods and Outcomes
- Cancer, Lipids, and Metabolism
- Epigenetics and DNA Methylation
- Cancer-related molecular mechanisms research
- Glutathione Transferases and Polymorphisms
- Metabolism, Diabetes, and Cancer
- Organ Transplantation Techniques and Outcomes
- Alkaline Phosphatase Research Studies
- RNA regulation and disease
University of Alberta
2013-2024
St Vincent's Hospital Sydney
2009
St. Vincent's Birmingham
2009
The palmitoylation of calnexin serves to enrich on the mitochondria-associated membrane (MAM). Given a lack information significance this finding, we have investigated how endoplasmic reticulum (ER)-internal sorting signal affects functions calnexin. Our results demonstrate that palmitoylated interacts with sarcoendoplasmic (SR) calcium transport ATPase (SERCA) 2b and interaction determines ER content regulation ER-mitochondria crosstalk. In contrast, non-palmitoylated oxidoreductase ERp57...
Endoplasmic reticulum (ER) homeostasis requires molecular regulators that tailor mitochondrial bioenergetics to the needs of protein folding. For instance, calnexin maintains mitochondria metabolism and mitochondria-ER contacts (MERCs) through reactive oxygen species (ROS) from NADPH oxidase 4 (NOX4). However, induction ER stress a quick rewiring adapt new energy needs. This machinery is not characterized. We now show oxidoreductase ERO1⍺ covalently interacts with kinase RNA-like (PERK) upon...
Progress in understanding the biology of protein fatty acylation has been impeded by lack rapid direct detection and identification methods. We first report that a synthetic omega-alkynyl-palmitate analog can be readily specifically incorporated into GAPDH or mitochondrial 3-hydroxyl-3-methylglutaryl-CoA synthase vitro reacted with an azido-biotin probe fluorogenic 3-azido-7-hydroxycoumarin using click chemistry for Western blotting flat bed fluorescence scanning. The acylated cysteine...
Huntington disease (HD) is a debilitating neurodegenerative characterized by the loss of motor control and cognitive ability that ultimately leads to death. It caused expansion polyglutamine tract in huntingtin (HTT) protein, which aggregation protein eventually cellular Both wild-type mutant form are highly regulated post-translational modifications including proteolysis, palmitoylation phosphorylation. We now demonstrate existence new modification HTT: addition 14 carbon fatty acid...
Control of Ca 2+ flow from the ER to mitochondria by calnexin balances output energy production pathways.
Myristoylation, the N-terminal modification of proteins with fatty acid myristate, is critical for membrane targeting and cell signaling. Because cancer cells often have increased N-myristoyltransferase (NMT) expression, NMTs were proposed as anti-cancer targets. To systematically investigate this, we performed robotic line screens discovered a marked sensitivity hematological lines, including B-cell lymphomas, to potent pan-NMT inhibitor PCLX-001. PCLX-001 treatment impacts global...
<p>Supplementary Figure S1: NMT1 and NMT2 have distinct expression patterns. is consistently expressed in ∼1 400 CCLE cell lines ∼11 000 TCGA patient samples. more variable entirely absent some cells. Expression measured by RNASeq as Log2(TPM+1). Data extracted from DepMap (CCLE, Public 23Q2) (via TCGABiolinks).</p>
<p>Supplementary Figure S3: NMT2 is associated with overall survival in the GSE37642 dataset: Patients dataset were separated into quartiles based on expression, and analyzed via Kaplan-Meier plots. Curves compared by log-rank test.</p>
<p>Supplementary Figure S5: MISS-54 Scores Categorized by AML Risk Category TCGA-LAML patients were separated into groups ELN2022 risk classification (A) or molecular category (B) and scores of compared one-way ANOVA. No significant differences found between groups.</p>
<p>Supplementary Figure S12: Matched viability assay for Resipher assays Cells were plated under identical conditions to in fig6D and measured by CellTitre BlueTM at 24 hours intervals (n = 2). No significant loss of was observed 72 (zelenirstat added hours).</p>
<p>Supplementary Figure S13: Zelenirstat reduces AMPKβ. AML cell lines were treated with zelenirstat for 48 hours, then stimulated 100 nL/mL SCF & FL, levels of AMPKβ analyzed by western blot. Band intensity was determined using Image Studio Lite, and each sample normalized to actin. Differences between treatment groups one-way ANOVA.</p>
<p>Supplementary Figure S4: Expression of NMT1 and NMT2 as it associates with AML risk factors. Patients were separated based on mutational status commonly used prognostic genes compared expression (A) (B). No significant differences found in displayed genes. into categories ELN2022 guidelines or TCGA-LAML molecular evaluated (C). found.</p>
<p>Supplementary Figure S14: Zelenirstat shows promising synergy with venetoclax. AML cell lines were screened for between zelenirstat and venetoclax using Horizon’s Combination Profiling platform. Isobolograms of (A) MV-4-11 (B) KG-1 combination therapy. Trendlines below additivity line indicate synergy. (C) Synergy scores in growth inhibition excess additive effects.</p>
<p>Supplementary Figure S2: Zelenirstat effectively inhibits myristoylation in AML cells. Cells pre-incubated with zelenirstat at indicated concentrations for 1 hour were metabolically labeled alkynyl-myristate analog. Incorporation of analog into proteins was measured by Western Blotting after click reaction azido-biotin using streptavidin-HRP/ECL.</p>
<p>Table of Materials</p>
<p>Supplementary Figure S6: Zelenirstat causes manageable weight loss in mice. Body of mice xenograft experiments fig3C (A) and fig3D (B). High-dose zelenirstat caused that was rapidly recovered after cessation treatment.</p>
<p>Figure S7: SFKs respond similarly to zelenirstat. AML cell lines were pre-treated with 1µM zelenirstat for 48 hours, then stimulated 100 ng/mL SCF & FL. Levels of indicated Src-family kinases analyzed by western blot.</p>
<p>Supplementary Figure S11: Zelenirstat reduces glycolytic rate in AML cells. MV-4-11 (A-B) and U937 (C-D) cells were incubated with zelenirstat for 48 hours, then extracellular acidification assessed using an Aglient Seahorse Glycolytic Rate Assay kit.</p>
<p>Figure S8: Zelenirstat disrupts signalling in AML. U937 and KG-1 cells were incubated with zelenirstat for 48 hours, stimulated 100 ng/mL FL & SCF, then lysed. Lysates probed HCK, phosphorylated SFKs, Stat5 by western blotting. Band intensity was determined using Image Studio Lite, each sample normalized to actin. Differences between treatment groups one-way ANOVA.</p>
<p>Supplementary Figure S9: Zelenirstat causes ER stress and Apoptosis in AML Cell Lines U937, KG-1 cell lines were incubated with zelenirstat for up to 72 hours, then lysed proteins analyzed by western blot. Representative blots of 3 biological replicates. Band intensity was determined using Image Studio Lite, each sample normalized actin. Differences between treatment groups one-way ANOVA.</p>
<p>Supplementary Figure S10: Zelenirstat reduces OCR in U937 cells were incubated with zelenirstat for 48 hours then analyzed using an Agilent Seahorse and Glycolytic Rate Assay kit (n = 5).</p>
<div>Abstract<p>Acute myeloid leukemia (AML) is a hematologic malignancy with limited treatment options and high likelihood of recurrence after chemotherapy. We studied N-myristoylation, the myristate modification proteins linked to survival signaling metabolism, as potential therapeutic target for AML. N-myristoylation catalyzed by two N-myristoyltransferases (NMT), NMT1 NMT2, varying expressions in AML cell lines patient samples. identified <i>NMT2</i> expression...
Abstract Background In humans, two ubiquitously expressed N-myristoyltransferases, NMT1 and NMT2, catalyze myristate transfer to proteins facilitate membrane targeting signaling. We investigated the expression of NMT s in numerous cancers found that NMT2 levels are dysregulated by epigenetic suppression, particularly so hematologic malignancies. This suggests pharmacological inhibition remaining could allow for selective killing these cells, sparing normal cells with both NMTs. Methods...
Myristoylation occurs cotranslationally on nascent proteins and post-translationally during apoptosis after caspase cleavages expose cryptic myristoylation sites. We demonstrate a drastic change in the myristoylated protein proteome apoptotic cells, likely as more substrates are revealed by caspases. show for first time that both N-myristoyltransferases (NMTs) 1 2 cleaved caspase-3- or -8-mediated cleavage of NMT1 at Asp-72 precedes NMT2 caspase-3 mainly Asp-25. The NMTs did not...