{"id":9633,"date":"2024-05-11T09:00:55","date_gmt":"2024-05-11T13:00:55","guid":{"rendered":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/?p=9633"},"modified":"2024-11-12T15:54:32","modified_gmt":"2024-11-12T20:54:32","slug":"c21-determination-of-ndm-4-metallo-%ce%b2-lactamase-structure-insights-on-antibiotic-resistance","status":"publish","type":"post","link":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/c21-determination-of-ndm-4-metallo-%ce%b2-lactamase-structure-insights-on-antibiotic-resistance\/","title":{"rendered":"C21: Determination of NDM-4 Metallo-\u03b2-lactamase Structure: Insights on Antibiotic Resistance"},"content":{"rendered":"\n

Through mutations and horizontal transfer, bacteria has been able to acquire different forms of antibiotic resistance. In particular, bacteria that produce \u03b2-lactamases as a result of mutated or acquired genes have the ability to break down the \u03b2-lactam ring structure in many common classes of antibiotics (penicillins, carbapenems, monobactams, and cephalosporins). Metallo-\u03b2-lactamases, enzymes that confer \u03b2-lactam antibiotic resistance, have shown an increased presence in the clinical setting and therefore have become a more significant issue. Structure determination for these enzymes helps future research regarding the search for inhibitors to be paired with antibiotics in new clinical regimens, disabling the enzyme\u2019s contribution to \u03b2-lactam antibiotic resistance. New Delhi Metallo-\u03b2-lactamase (NDMs), a member of the B1 subclass of metallo-\u03b2-lactamases, has been gaining recognition due to its prevalence in the clinic, yet the field requires more research on its structure. This study focuses on the variant NDM-4. Activity assays were performed with NDM-4 to determine activity prior to any modifications. A new NDM-4 construct, NDM-4_6-Aldolase, was developed and was studied for the purpose of using cryo-EM to solve the structure of NDM-4. The tetramer of aldolase (PDB code 1ZAH, Fructose-1,6-bisphosphate aldolase) used in these constructs was chosen for its large size and symmetrical tetrameric structure. These constructs fused NDM-4 and aldolase to produce a construct amenable for Cryo-EM. Activity assays and a cryo-EM structure were successfully obtained. NDM-4_6-Aldolase exhibited a best-fit Km of 128 \u00b5M. Cryo-EM will be performed to resolve the structure of this new construct. Future work with dipicolinic acid (DPA) as an inhibitor of NDM-4_6-Aldolase is planned.<\/p>\n\n\n\n

Authors: S. Kei Brown, Nastia Evstifeeva, & Richard C. Page, Ph.D.<\/p>\n\n\n\n

Advisor(s): Richard Page, Chemistry & Biochemistry<\/p>\n\n\n\n

Nastia Evstifeeva, Chemistry & Biochemistry<\/p>\n\n\n\n

\"\"<\/figure>\n","protected":false},"excerpt":{"rendered":"

Through mutations and horizontal transfer, bacteria has been able to acquire different forms of antibiotic resistance. In particular, bacteria that produce \u03b2-lactamases as a result of mutated or acquired genes have the ability to break down the \u03b2-lactam ring structure in many common classes of antibiotics (penicillins, carbapenems, monobactams, and cephalosporins). Metallo-\u03b2-lactamases, enzymes that confer […]<\/p>\n","protected":false},"author":301,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_bbp_topic_count":0,"_bbp_reply_count":0,"_bbp_total_topic_count":0,"_bbp_total_reply_count":0,"_bbp_voice_count":0,"_bbp_anonymous_reply_count":0,"_bbp_topic_count_hidden":0,"_bbp_reply_count_hidden":0,"_bbp_forum_subforum_count":0,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2},"jetpack_post_was_ever_published":false},"categories":[1589,1,1605,27],"tags":[1029,652,89,1697],"class_list":["post-9633","post","type-post","status-publish","format-standard","hentry","category-2024-posters","category-projects","category-c21-30-2024-posters","category-chemistry-biochemistry","tag-antibiotic-resistance","tag-biochemistry","tag-stem","tag-structural-biology"],"aioseo_notices":[],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/pbRpLx-2vn","jetpack_likes_enabled":true,"_links":{"self":[{"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/posts\/9633","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/users\/301"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/comments?post=9633"}],"version-history":[{"count":0,"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/posts\/9633\/revisions"}],"wp:attachment":[{"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/media?parent=9633"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/categories?post=9633"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/wp-json\/wp\/v2\/tags?post=9633"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}