{"id":866,"date":"2024-09-05T10:14:37","date_gmt":"2024-09-05T10:14:37","guid":{"rendered":"http:\/\/tezcan.ucsd.edu\/?page_id=866"},"modified":"2024-09-19T16:44:01","modified_gmt":"2024-09-19T23:44:01","slug":"nitrogenase-structure-and-function","status":"publish","type":"page","link":"http:\/\/tezcan.ucsd.edu\/index.php\/nitrogenase-structure-and-function\/","title":{"rendered":"Nitrogenase Structure and Function"},"content":{"rendered":"\n<div class=\"wp-block-columns \\&quot;wp-block-columns\\&quot; is-layout-flex wp-container-core-columns-is-layout-1 wp-block-columns-is-layout-flex\" style=\"padding-right:150px;padding-left:150px\">\n<div class=\"wp-block-column \\&quot;wp-block-column\\&quot; is-layout-flow wp-block-column-is-layout-flow\">\n<div class=\"wp-swiper\"><div class=\"wp-swiper__wrapper\"><div class=\"swiper-container swiper\" data-swiper=\"{&quot;data-slidesperview&quot;:&quot;1&quot;,&quot;data-navigation&quot;:true,&quot;data-pagination&quot;:true,&quot;data-autoplay&quot;:false,&quot;data-disableOnInteraction&quot;:true,&quot;data-pauseOnMouseEnter&quot;:false,&quot;data-delay&quot;:3000,&quot;data-speed&quot;:500,&quot;data-loop&quot;:false,&quot;data-effect&quot;:&quot;slide&quot;,&quot;data-direction&quot;:&quot;horizontal&quot;,&quot;data-freemode&quot;:false,&quot;data-autoheight&quot;:true,&quot;data-spacebetween&quot;:0,&quot;data-mousewheel&quot;:false,&quot;data-releaseonedges&quot;:false,&quot;data-paginationtype&quot;:&quot;bullets&quot;}\" data-slidesperview=\"1\" data-navigation=\"true\" data-pagination=\"true\" data-autoplay=\"false\" data-disableoninteraction=\"true\" data-pauseonmouseenter=\"false\" data-delay=\"3000\" data-speed=\"500\" data-loop=\"false\" data-effect=\"slide\" data-direction=\"horizontal\" data-freemode=\"false\" data-autoheight=\"true\" data-spacebetween=\"0\" data-mousewheel=\"false\" data-releaseonedges=\"false\" data-paginationtype=\"bullets\"><div class=\"swiper-wrapper\">\n<div data-tab=\"slide-1\" class=\"wp-swiper__slide swiper-slide\"><div class=\"wp-swiper__overlay-color\" style=\"background-color:rgba(0, 0, 0, 0)\"><\/div><div class=\"wp-swiper__slide-content\">\n<figure style=\"padding-right:72px;padding-left:72px\" class=\"wp-block-video\"><video controls src=\"http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/FePApproach_3.mp4\"><\/video><\/figure>\n\n\n\n<p class=\"has-text-align-center has-medium-font-size\">Nitrogenase docking geometries determined by X-ray crystallography and cryoEM<\/p>\n\n\n\n<div style=\"height:40px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n<\/div><\/div>\n\n\n\n<div data-tab=\"slide-2\" class=\"wp-swiper__slide swiper-slide\"><div class=\"wp-swiper__overlay-color\" style=\"background-color:rgba(0, 0, 0, 0)\"><\/div><div class=\"wp-swiper__slide-content\">\n<figure style=\"padding-right:72px;padding-left:72px\" class=\"wp-block-video\"><video controls src=\"http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/ATPATP_18.mp4\"><\/video><\/figure>\n\n\n\n<p class=\"has-text-align-center has-medium-font-size\">Nitrogenase complexes captured under catalytic turnover with cryoEM<\/p>\n\n\n\n<div style=\"height:var(--wp--preset--spacing--small)\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n<\/div><\/div>\n\n\n\n<div data-tab=\"slide-3\" class=\"wp-swiper__slide swiper-slide\"><div class=\"wp-swiper__overlay-color\" style=\"background-color:rgba(0, 0, 0, 0)\"><\/div><div class=\"wp-swiper__slide-content\">\n<figure class=\"wp-block-video\"><video controls src=\"http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/firstRotateDens_2.mp4\"><\/video><\/figure>\n\n\n\n<p class=\"has-text-align-center has-medium-font-size\">FeSII-nitrogenase complex (2.27 \u00c5, C2) structurally determined by cryoEM<\/p>\n\n\n\n<div style=\"height:var(--wp--preset--spacing--small)\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n<\/div><\/div>\n<\/div><\/div><div class=\"wp_swiper__navigation\"><div class=\"wp_swiper__navigation-container\"><div class=\"swiper-button-prev \"><\/div><div class=\"swiper-button-next \"><\/div><\/div><\/div><div class=\"swiper-pagination\"><\/div><\/div><\/div>\n<\/div>\n<\/div>\n\n\n\n<p>The catalytic reduction of atmospheric dinitrogen into ammonia (i.e., reductive nitrogen fixation) is a key process for the biosynthesis of life\u2019s building blocks like amino and nucleic acids. Approximately, half of fixed nitrogen in our biosphere is provided by the industrial Haber-Bosch process and the other half by nitrogenases produced by diazotropic bacteria. Despite its remarkable efficiency, the Haber-Bosch process requires extreme conditions (&gt;400 <sup>o<\/sup>C, &gt;250 atm H<sub>2<\/sub>\/N<sub>2<\/sub>),&nbsp; is highly energy intensive (accounting for ~2% of all global energy consumption), and a major contributor to greenhouse gas emissions. In contrast, nitrogenase catalysis occurs at ambient conditions of bacterial growth, albeit at the expense of a large number of ATP molecules required to control the flow of electrons and protons to enable nitrogen reduction by nitrogenase.<sup>1<\/sup><\/p>\n\n\n\n<p>Although nitrogenase has been studied extensively for over six decades, the key details of its catalytic mechanism, biosynthesis, and cellular activities are yet to be elucidated: (a) what is the mechanism of nitrogen reduction by the catalytic cofactor (FeMo-co) of nitrogenase? (b) how does ATP hydrolysis control\/gate the flow of multiple electrons and protons to FeMoco? (c) how are the complex cofactors of nitrogenase synthesized and incorporated into nitrogenase, (d) how does nitrogenase interact with other cellular components? For answering the first two questions, our group previously developed strategies for light-driven systems for ATP-independent nitrogenase catalysis<sup>2,3<\/sup> and elucidated key roles of dynamic\/multi-modal protein-protein interactions in ATP-mediated electron transfer and redox-dependent conformational gating events.<sup>4-7<\/sup> Yet, these studies also highlighted the need for new methods for monitoring the structure of nitrogenase during catalytic turnover. Toward this end, we recently joined forces with the Herzik Group at UCSD Chemistry\/Biochemistry to capture the structural dynamics of nitrogenase in action by anaerobic cryoEM and detected new conformational states and cofactor dynamics of nitrogenase that could not be visualized by other methods.<sup>8<\/sup> Using these findings as a springboard, the current goals of the nitrogenase sub-group are to visualize catalytic reaction intermediates of nitrogenase at atomic resolution, understand the nature of ATP-directed electron and proton transfer, and characterize the dynamics interactions of nitrogenase with other protein components involved in its biosynthesis and its cellular functions.<\/p>\n\n\n\n<div style=\"height:var(--wp--preset--spacing--x-large)\" aria-hidden=\"true\" class=\"wp-block-spacer \\&quot;wp-block-spacer\\&quot;\"><\/div>\n\n\n\n<p class=\"has-text-align-center \\&quot;has-text-align-center\\&quot;\">&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8211; Selected Publications &#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8212;&#8211;<\/p>\n\n\n\n<div style=\"height:var(--wp--preset--spacing--medium)\" aria-hidden=\"true\" class=\"wp-block-spacer \\&quot;wp-block-spacer\\&quot;\"><\/div>\n\n\n\n<p>1. Rutledge, H. L. &amp; Tezcan, F. A. Electron transfer in nitrogenase. <em>Chem. Rev.<\/em> <strong>120<\/strong>, 5158-5193 (2020).<\/p>\n\n\n\n<p>2. Roth, L. E., Nguyen, J. C. &amp; Tezcan, F. A. ATP- and iron-protein-independent activation of nitrogenase catalysis by light. <em>J. Am. Chem. Soc. <\/em><strong>132<\/strong>, 13672-13674 (2010).<\/p>\n\n\n\n<p>3. Roth, L. E. &amp; Tezcan, F. A. ATP-Uncoupled, Six-Electron Photoreduction of Hydrogen Cyanide to Methane by the Molybdenum-Iron Protein. <em>J. Am. Chem. Soc.<\/em> <strong>134<\/strong>, 8416-8419 (2012).<\/p>\n\n\n\n<p>4. Owens, C. P., Katz, F. E., Carter, C. H., Luca, M. A. &amp; Tezcan, F. A. Evidence for functionally relevant encounter complexes in nitrogenase catalysis. <em>J. Am. Chem. Soc.<\/em> <strong>137<\/strong>, 12704-12712 (2015).<\/p>\n\n\n\n<p>5. Owens, C. P., Katz, F. E. H., Carter, C. H., Oswald, V. F. &amp; Tezcan, F. A. Tyrosine-Coordinated P-Cluster in G. diazotrophicus Nitrogenase: Evidence for the Importance of O-Based Ligands in Conformationally Gated Electron Transfer. <em>J. Am. Chem. Soc.<\/em> <strong>138<\/strong>, 10124-10127 (2016).<\/p>\n\n\n\n<p>6. Rutledge, H. L., Field, M. J., Rittle, J., Green, M. T. &amp; Tezcan, F. A. Role of Serine Coordination in the Structural and Functional Protection of the Nitrogenase P-Cluster. <em>J. Am. Chem. Soc.<\/em> <strong>144<\/strong>, 22101-22112 (2022).<\/p>\n\n\n\n<p>7. Rutledge, H. L.<em> et al.<\/em> Redox-Dependent Metastability of the Nitrogenase P-Cluster. <em>J. Am. Chem. Soc.<\/em> <strong>141<\/strong>, 10091-10098 (2019).<\/p>\n\n\n\n<p>8. Rutledge, H. L., Cook, B. D., Nguyen, H. P. M., Herzik, M. A. &amp; Tezcan, F. A. Structures of the nitrogenase complex prepared under catalytic turnover conditions. <em>Science<\/em> <strong>377<\/strong>, 865-869 (2022).<\/p>\n\n\n\n<div style=\"height:var(--wp--preset--spacing--medium)\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<div class=\"wp-block-buttons is-content-justification-center is-layout-flex wp-container-core-buttons-is-layout-1 wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button is-style-outline is-style-outline--16c846e6b1a0909c2e5ae2f0a7d7bd93\"><a class=\"wp-block-button__link wp-element-button\" href=\"http:\/\/tezcan.ucsd.edu\/index.php\/research-2\/\">Go Back<\/a><\/div>\n<\/div>\n\n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>The catalytic reduction of atmospheric dinitrogen into ammonia (i.e., reductive nitrogen fixation) is a key process for the biosynthesis of life\u2019s building blocks like amino and nucleic acids. Approximately, half of fixed nitrogen in our biosphere is provided by the industrial Haber-Bosch process and the other half by nitrogenases produced by diazotropic bacteria. Despite its &hellip; <\/p>\n<p class=\"link-more\"><a href=\"http:\/\/tezcan.ucsd.edu\/index.php\/nitrogenase-structure-and-function\/\" class=\"more-link\">Read more<span class=\"screen-reader-text\"> &#8220;Nitrogenase Structure and Function&#8221;<\/span><\/a><\/p>\n","protected":false},"author":2,"featured_media":1228,"parent":0,"menu_order":2,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-866","page","type-page","status-publish","has-post-thumbnail","hentry"],"featured_media_urls":{"thumbnail":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",150,75,false],"medium":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",300,150,false],"medium_large":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",768,384,false],"large":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",950,475,false],"1536x1536":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",1536,768,false],"2048x2048":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",2000,1000,false],"inspiro-featured-image":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",2000,1000,false],"inspiro-loop":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",640,320,false],"inspiro-loop@2x":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",1280,640,false],"portfolio_item-thumbnail":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",600,300,false],"portfolio_item-thumbnail@2x":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",1200,600,false],"portfolio_item-masonry":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",600,300,false],"portfolio_item-masonry@2x":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",1200,600,false],"portfolio_item-thumbnail_cinema":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",670,335,false],"portfolio_item-thumbnail_portrait":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",600,300,false],"portfolio_item-thumbnail_portrait@2x":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",1200,600,false],"portfolio_item-thumbnail_square":["http:\/\/tezcan.ucsd.edu\/wp-content\/uploads\/2024\/09\/DSCF5645-2.jpg",800,400,false]},"_links":{"self":[{"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/pages\/866"}],"collection":[{"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/comments?post=866"}],"version-history":[{"count":32,"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/pages\/866\/revisions"}],"predecessor-version":[{"id":1353,"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/pages\/866\/revisions\/1353"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/media\/1228"}],"wp:attachment":[{"href":"http:\/\/tezcan.ucsd.edu\/index.php\/wp-json\/wp\/v2\/media?parent=866"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}