{"id":556,"date":"2020-10-09T12:16:48","date_gmt":"2020-10-09T10:16:48","guid":{"rendered":"https:\/\/sites.unimi.it\/apisgroup\/?p=556"},"modified":"2022-01-21T13:00:14","modified_gmt":"2022-01-21T12:00:14","slug":"thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability","status":"publish","type":"post","link":"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/","title":{"rendered":"Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC\u2013LptA Interaction and Decreasing LptA Stability"},"content":{"rendered":"\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"677\" src=\"https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-1024x677.jpg\" alt=\"\" class=\"wp-image-558\" srcset=\"https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-1024x677.jpg 1024w, https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-300x198.jpg 300w, https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-768x508.jpg 768w, https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-1536x1015.jpg 1536w, https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina.jpg 1950w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p class=\"has-text-align-left wp-block-paragraph\">The outer membrane (OM) of Gram-negative bacteria is a highly selective permeability barrier due to its asymmetric structure with lipopolysaccharide (LPS) in the outer leaflet. In Escherichia coli, LPS is transported to the cell surface by the LPS transport (Lpt) system composed of seven essential proteins forming a transenvelope bridge. Transport is powered by the ABC transporter LptB2FGC, which extracts LPS from the inner membrane (IM) and transfers it, through LptC protein, to the periplasmic protein LptA. Then, LptA delivers LPS to the OM LptDE translocon for final assembly at the cell surface. The Lpt protein machinery operates as a single device, since depletion of any component leads to the accumulation of a modified LPS decorated with repeating units of colanic acid at the IM outer leaflet. Moreover, correct machine assembly is essential for LPS transit and disruption of the Lpt complex results in LptA degradation. Due to its vital role in cell physiology, the Lpt system represents a good target for antimicrobial drugs. Thanatin is a naturally occurring antimicrobial peptide reported to cause defects in membrane assembly and demonstrated in vitro to bind to the N-terminal b-strand of LptA. Since this region is involved in both LptA dimerization and interaction with LptC, we wanted to elucidate the mechanism of inhibition of thanatin and discriminate whether its antibacterial effect is exerted by the disruption of the interaction of LptA with itself or with LptC. For this purpose, we here implemented the Bacterial Adenylate Cyclase Two-<br>Hybrid (BACTH) system to probe in vivo the Lpt interactome in the periplasm. With this system, we found that thanatin targets both LptC\u2013LptA and LptA\u2013LptA interactions, with a greater inhibitory effect on the former. We confirmed in vitro the disruption of LptC\u2013LptA interaction using two different biophysical techniques. Finally, we observed that in<br>cells treated with thanatin, LptA undergoes degradation and LPS decorated with colanic acid accumulates. These data further support inhibition or disruption of Lpt complex assembly as the main killing mechanism of thanatin against Gram-negative bacteria.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"doi: 10.3389\/fmicb.2020.00909\">doi: 10.3389\/fmicb.2020.00909<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>The outer membrane (OM) of Gram-negative bacteria is a highly selective permeability barrier due to its asymmetric structure with lipopolysaccharide (LPS) in the outer leaflet. In Escherichia coli, LPS is transported to the cell&#46;&#46;&#46;<\/p>\n","protected":false},"author":1,"featured_media":559,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[6],"tags":[29,28],"class_list":["post-556","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-application","tag-gram-negative-bacteria","tag-thanatin"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v26.8 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC\u2013LptA Interaction and Decreasing LptA Stability - APIS group<\/title>\n<meta name=\"robots\" content=\"noindex, follow\" \/>\n<meta property=\"og:locale\" content=\"it_IT\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC\u2013LptA Interaction and Decreasing LptA Stability - APIS group\" \/>\n<meta property=\"og:description\" content=\"The outer membrane (OM) of Gram-negative bacteria is a highly selective permeability barrier due to its asymmetric structure with lipopolysaccharide (LPS) in the outer leaflet. In Escherichia coli, LPS is transported to the cell&#046;&#046;&#046;\" \/>\n<meta property=\"og:url\" content=\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/\" \/>\n<meta property=\"og:site_name\" content=\"APIS group\" \/>\n<meta property=\"article:published_time\" content=\"2020-10-09T10:16:48+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2022-01-21T12:00:14+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-1.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1950\" \/>\n\t<meta property=\"og:image:height\" content=\"1289\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"emanuela erba\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Scritto da\" \/>\n\t<meta name=\"twitter:data1\" content=\"emanuela erba\" \/>\n\t<meta name=\"twitter:label2\" content=\"Tempo di lettura stimato\" \/>\n\t<meta name=\"twitter:data2\" content=\"2 minuti\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/#article\",\"isPartOf\":{\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/\"},\"author\":{\"name\":\"emanuela erba\",\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/#\/schema\/person\/983a9690b88e39642466bb60ddc30c84\"},\"headline\":\"Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC\u2013LptA Interaction and Decreasing LptA Stability\",\"datePublished\":\"2020-10-09T10:16:48+00:00\",\"dateModified\":\"2022-01-21T12:00:14+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/\"},\"wordCount\":352,\"publisher\":{\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/#organization\"},\"image\":{\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/#primaryimage\"},\"thumbnailUrl\":\"https:\/\/sites.unimi.it\/apisgroup\/wp-content\/uploads\/2020\/10\/tanatina-1.jpg\",\"keywords\":[\"Gram-negative bacteria\",\"thanatin\"],\"articleSection\":[\"application\"],\"inLanguage\":\"it-IT\"},{\"@type\":\"WebPage\",\"@id\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/\",\"url\":\"https:\/\/sites.unimi.it\/apisgroup\/2020\/10\/09\/thanatin-impairs-lipopolysaccharide-transport-complex-assembly-by-targeting-lptc-lpta-interaction-and-decreasing-lpta-stability\/\",\"name\":\"Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC\u2013LptA Interaction and Decreasing LptA Stability - 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