{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:45:27Z","timestamp":1760402727271,"version":"build-2065373602"},"reference-count":47,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,4,2]],"date-time":"2021-04-02T00:00:00Z","timestamp":1617321600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BIA-BQM\/30421\/2017","PD\/BD\/128261\/2016"],"award-info":[{"award-number":["PTDC\/BIA-BQM\/30421\/2017","PD\/BD\/128261\/2016"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010665","name":"H2020 Marie Sk\u0142odowska-Curie Actions","doi-asserted-by":"publisher","award":["823780"],"award-info":[{"award-number":["823780"]}],"id":[{"id":"10.13039\/100010665","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Instruct-ULTRA","award":["731005"],"award-info":[{"award-number":["731005"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM132120","GM116799"],"award-info":[{"award-number":["GM132120","GM116799"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Processes"],"abstract":"<jats:p>Studies on membrane proteins can help to develop new drug targets and treatments for a variety of diseases. However, membrane proteins continue to be among the most challenging targets in structural biology. This uphill endeavor can be even harder for membrane proteins from Mycobacterium species, which are notoriously difficult to express in heterologous systems. Arabinofuranosyltransferases are involved in mycobacterial cell wall synthesis and thus potential targets for antituberculosis drugs. A set of 96 mycobacterial genes coding for Arabinofuranosyltransferases was selected, of which 17 were successfully expressed in E. coli and purified by metal-affinity chromatography. We herein present an efficient high-throughput strategy to screen in microplates a large number of targets from Mycobacteria and select the best conditions for large-scale protein production to pursue functional and structural studies. This methodology can be applied to other targets, is cost and time effective and can be implemented in common laboratories.<\/jats:p>","DOI":"10.3390\/pr9040629","type":"journal-article","created":{"date-parts":[[2021,4,2]],"date-time":"2021-04-02T10:34:09Z","timestamp":1617359649000},"page":"629","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["High Throughput Expression Screening of Arabinofuranosyltransferases from Mycobacteria"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0829-9054","authenticated-orcid":false,"given":"Jos\u00e9","family":"Rodrigues","sequence":"first","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5552-4205","authenticated-orcid":false,"given":"Vanessa T.","family":"Almeida","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"}]},{"given":"Ana L.","family":"Ros\u00e1rio","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6656-6320","authenticated-orcid":false,"given":"Yong Zi","family":"Tan","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY 10032, USA"},{"name":"National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY 10027, USA"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0130-8739","authenticated-orcid":false,"given":"Brian","family":"Kloss","sequence":"additional","affiliation":[{"name":"Center on Membrane Protein Production and Analysis, New York Structural Biology Center, New York, NY 10027, USA"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3293-2200","authenticated-orcid":false,"given":"Filippo","family":"Mancia","sequence":"additional","affiliation":[{"name":"Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY 10032, USA"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8419-5420","authenticated-orcid":false,"given":"Margarida","family":"Archer","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1002\/pro.5560070420","article-title":"Genome-wide analysis of integral membrane proteins from eubacterial, archaean, and eukaryotic organisms","volume":"7","author":"Wallin","year":"1998","journal-title":"Protein Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1006\/jmbi.2000.4315","article-title":"Predicting transmembrane protein topology with a hidden markov model: Application to complete genomes11Edited by F","volume":"305","author":"Krogh","year":"2001","journal-title":"Cohen. 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