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<article><div class="banner banner--gray banner--compact" style="--banner-background-image: url('/sites/default/files/media/image/2021-03/2015.jpg');--banner-background-focus: 50% 50%"><div class="banner__wrapper"><div class="banner__container"><h2 class="banner__heading">NIH Research Festival</h2><div class="banner__content"><p><strong><time datetime="2015-09-16">September 16</time> – <time datetime="2015-09-18">18, 2015</time></strong></p></div></div></div></div></article>
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<div class="section"><div data-drupal-messages-fallback class="hidden"></div></div><main role="main"><div id="main-content" tabindex="-1"></div><div class="section"><div class="content-sidebar"><div class="content-sidebar__main"><div class="content-sidebar__main-content"><div id="block-mainpagecontent-2"><article><h1>Identification of a novel inhibitor of 5-Lipoxygenase</h1><div class="schedule"><div class="schedule__label"><div class="schedule__label-text"><time datetime="2015-09-16">Wednesday, September 16, 2015</time>
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– Poster Session I
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</div></div><div class="schedule__content"><div class="schedule__item"><div class="schedule__time">3:30 – 5:00 p.m.</div><div class="schedule__description"><p><strong>FAES Terrace</strong></p></div><div class="schedule__description"><p><strong>NEI</strong></p></div><div class="schedule__description"><p><strong>MOLBIO-31</strong></p></div></div></div></div><h2>Authors</h2><ul><li>P Subramanian</li><li>E Mendez</li><li>SP Becerra</li></ul><h2>Abstract</h2><p>Arachidonate 5-lipoxygenase (5-LOX) is a member of the lipoxygenase family of enzymes that plays a key role in arachidonic acid metabolism. Arachidonic acid, upon oxygenation by 5-LOX, forms 5-hydroperoxyeicosatetraenoic acid (5-HPETE), which is further converted into proinflammatory leukotrienes such as LTB4 that can have detrimental biological effects. The PNPLA2 gene codes for PEDF-R, a lipase involved in cell survival and lipid metabolism, found in the RPE and retina. Peptides derived from the longest loop region (L159-M325) of PEDF-R were screened for the effect on soybean lipoxygenase-V (LOX-V) in vitro. We identified PEDF-R peptides E5b (I193–L232) and P1 (T210-L249) with binding affinity for LOX-V that inhibited its enzymatic activity. The LOX-V enzymatic inhibition was attenuated when PEDF-R peptides were captured specifically with a P1 binding protein, fluorescein-conjugated PEDF (Fl-PEDF). These peptides efficiently protected human retinal pigment epithelial ARPE-19 and pig primary RPE cells from oxidative stress. We investigated whether binding and inhibiting mammalian 5-LOX by these peptides are critical to protect RPE cells during oxidative stress. Peptide affinity chromatography showed that human recombinant 5-LOX binds to P1 via ionic interactions. Pull-down assays revealed that full length PEDF-R interacts with 5-LOX in a dose-response fashion. Interestingly, PNPLA2 expression decreased with oxidative stress with no effect on 5-LOX expression. Moreover, addition of P1 and PEDF-R over-expression reduced of the levels of the 5-LOX product LTB4 in ARPE-19 cells under oxidative stress. Our results support that PEDF-R is a novel inhibitor of 5-LOX.</p><p><strong>Scientific Focus Area</strong>: Molecular Biology and Biochemistry</p><p class="updated"><small>This page was last updated on Friday, March 26, 2021</small></p></article>
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