{"id":1585,"date":"2020-01-13T01:20:00","date_gmt":"2020-01-13T00:20:00","guid":{"rendered":"https:\/\/www.nanoworld.com\/blog\/?p=1585"},"modified":"2023-04-18T12:59:24","modified_gmt":"2023-04-18T11:59:24","slug":"real-time-dynamics-of-gating-related-conformational-changes-in-cora","status":"publish","type":"post","link":"https:\/\/www.nanoworld.com\/blog\/real-time-dynamics-of-gating-related-conformational-changes-in-cora\/","title":{"rendered":"Real time dynamics of Gating-Related conformational changes in CorA"},"content":{"rendered":"\n

Magnesium (Mg2+<\/sup>) is a key\ndivalent cation in biology. It regulates\nand maintains numerous, physiological functions such as nucleic acid stability,\nmuscle contraction, heart rate and vascular tone, neurotransmitter release, and\nserves as cofactor in a myriad of enzymatic reactions. Most\nimportantly, it coordinates with ATP, and is thus crucial for energy production\nin mitochondria.*<\/p>\n\n\n\n

In order to\nstore Mg2+<\/sup> in the mitochondrial lumen it is imported via Mrs2 and\nAlr2 ion channels that are closely related to CorA, the main Mg2+-importer in\nbacteria. Although these Mg2+-transport proteins do not show much sequence\nconservation, they all share two trans-membrane domains (TMDs) with the\nsignature motif Glycine-Methionine-Asparagine (GMN) at the extracellular loop.*<\/p>\n\n\n\n

CorA, a divalent-selective channel in the\nmetal ion transport superfamily, is the major Mg2+<\/sup>-influx pathway in\nprokaryotes. CorA structures in closed (Mg2+<\/sup>-bound), and open (Mg2+<\/sup>-free)\nstates, together with functional data showed that Mg2+<\/sup>-influx\ninhibits further Mg2+<\/sup>-uptake completing a regulatory feedback loop.\nWhile the closed state structure is a symmetric pentamer, the open state\ndisplayed unexpected asymmetric architectures.*<\/p>\n\n\n\n

In the\narticle \u201cReal time dynamics of Gating-Related conformational changes in CorA\u201d Martina\nRangl, Nicolaus Schmandt, Eduardo Perozo and Simon Scheuring used high-speed atomic force microscopy (HS-AFM), to explore\nthe Mg2+<\/sup>-dependent gating transition of single CorA channels: HS-AFM\nmovies during Mg2+<\/sup>-depletion experiments revealed the channel\u2019s\ntransition from a stable Mg2+<\/sup>-bound state over a highly mobile and\ndynamic state with fluctuating subunits to asymmetric structures with varying\ndegree of protrusion heights from the membrane.*<\/p>\n\n\n\n

Their data shows that at Mg2+<\/sup>-concentration below Kd, CorA\nadopts a dynamic (putatively open) state of multiple conformations that imply\nstructural rearrangements through hinge-bending in TM1. They also\ndiscuss how these structural dynamics define the functional behavior of this\nligand-dependent channel.*<\/p>\n\n\n\n

All Atomic Force Microscopy experiments described in the article were performed using NanoWorld Ultra-Short Cantilevers<\/a> USC-F1.2-k0.15<\/a> for high-speed Atomic Force Microscopy ( HS-AFM ). Videos of CorA membranes were recorded with imaging rates of ~1\u20132 frames s\u22121 and at a resolution of 0.5 nm pixel\u22121.<\/p>\n\n\n\n

\"\"
Figure 1 from \u201cReal time dynamics of Gating-Related conformational changes in CorA<\/em>\u201d:
Sample morphology of CorA reconstitutions for HS-AFM.
 
(a) HS-AFM overview topograph of densely packed CorA in a POPC\/POPG (3:1) lipid bilayer exposing the periplasmic side and a loosely packed protein area with diffusing molecules exposing the intracellular face (full color scale: 20 nm). Left: Height histogram of the HS-AFM image with two peaks representative of the mica and the CorA surface (\u2206Height (peak-peak): 12 nm (20,500 height values)). The dashed line indicates the position of the cross-section analysis shown in (b). (b) Profile of the membrane shown in a), including a cartoon (top) of the membrane in side view. The height profile (~12 nm) corresponds well to the all-image height analysis (a, left) and the CorA structure (Matthies et al., 2016). (c) High-resolution image (top) and cross-section analysis along dashed line (bottom) of the periplasmic face. The height and dimension of the periplasmic face is in good agreement with the structure (left), and the periodicity (~14 nm, n = 40) corresponds well with the diameter of the intracellular face spacing the molecules on the other side of the membrane (full color scale: 2 nm). (d) HS-AFM image of densely packed CorA embedded in a DOPC\/DOPE\/DOPS (4:5:1) membrane. This reconstitution resulted in two stacked membrane layers, both exposing the CorA intracellular face. The dashed line indicates the position of the cross-section analysis shown in (e). Left: Height histogram of the HS-AFM image with two peaks at ~12 nm and ~17 nm (32,500 height values), corresponding to the proteins in two stacked membranes (full color scale: 20 nm). (e) Section profile of the membrane shown in d), including a cartoon (top) of the membrane in side view. (f) High-resolution view and cross-section analysis along dashed line (bottom) of the CorA intracellular face revealing the individual subunits of the pentamers (full color scale: 3 nm). Inset: 5-fold symmetrized average of CorA. The dimensions of CorA observed with HS-AFM are in good agreement with the structure (left: PDB 3JCF). The structures in (c) and (f) are shown in ribbon (top) and surface (bottom) representations, respectively. <\/figcaption><\/figure>\n\n\n\n

*Martina Rangl, Nicolaus Schmandt, Eduardo Perozo, and Simon Scheuring
Real time dynamics of Gating-Related conformational changes in CorA<\/strong>
eLife. 2019; 8: e47322
DOI: 10.7554\/eLife.47322<\/p>\n\n\n\n

Please follow this external link to read the full article: https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6927688\/<\/a><\/p>\n\n\n\n

Open Access: The article \u201cReal time dynamics of Gating-Related conformational changes in CorA\u201d by Martina Rangl, Nicolaus Schmandt, Eduardo Perozo and Simon Scheuring is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article\u2019s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article\u2019s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http:\/\/creativecommons.org\/licenses\/by\/4.0\/<\/a>. <\/p>\n","protected":false},"excerpt":{"rendered":"

Magnesium (Mg2+) is a key divalent cation in biology. It regulates and maintains numerous, physiological functions such as nucleic acid stability, muscle contraction, heart rate and vascular tone, neurotransmitter release, and serves as cofactor in a myriad of enzymatic reactions. Most importantly, it coordinates with ATP, and is thus crucial for energy production in mitochondria.* … Continue reading Real time dynamics of Gating-Related conformational changes in CorA<\/span><\/a><\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[3],"tags":[93,48,97,50,25,24,134,47,250,52,26,53,124,51,121,392,393,394,395,396],"class_list":{"0":"post-1585","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"hentry","6":"category-news","7":"tag-biology","8":"tag-biology-afm-probes","9":"tag-cell-biology","10":"tag-high-speed-afm","11":"tag-high-speed-scanning","12":"tag-hs-afm","13":"tag-life-science","14":"tag-life-science-afm-probes","15":"tag-molecular-biology","16":"tag-ultra-short-afm-cantilevers","17":"tag-ultrafast-scanning","18":"tag-usc","19":"tag-usc-f1-2-k0-15","20":"tag-video-rate-afm","21":"tag-video-rate-atomic-force-microscopy","22":"tag-afm","25":"tag-395","26":"tag-396"},"_links":{"self":[{"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/posts\/1585","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/comments?post=1585"}],"version-history":[{"count":5,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/posts\/1585\/revisions"}],"predecessor-version":[{"id":1777,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/posts\/1585\/revisions\/1777"}],"wp:attachment":[{"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/media?parent=1585"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/categories?post=1585"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.nanoworld.com\/blog\/wp-json\/wp\/v2\/tags?post=1585"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}