{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T06:56:57Z","timestamp":1792133817009,"version":"4.4.0"},"reference-count":46,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,11,2]],"date-time":"2018-11-02T00:00:00Z","timestamp":1541116800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2018,11,2]],"date-time":"2018-11-02T00:00:00Z","timestamp":1541116800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Commun Biol"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Nitrilases are oligomeric, helix-forming enzymes from plants, fungi and bacteria that are involved in the metabolism of various natural and artificial nitriles. These biotechnologically important enzymes are often specific for certain substrates, but directed attempts at modifying their substrate specificities by exchanging binding pocket residues have been largely unsuccessful. Thus, the basis for their selectivity is still unknown. Here we show, based on work with two highly similar nitrilases from the plant <jats:italic>Capsella rubella<\/jats:italic>, that modifying nitrilase helical twist, either by exchanging an interface residue or by imposing a different twist, without altering any binding pocket residues, changes substrate preference. We reveal that helical twist and substrate size correlate and when binding pocket residues are exchanged between two nitrilases that show the same twist but different specificities, their specificities change. Based on these findings we propose that helical twist influences the overall size of the binding pocket.<\/jats:p>","DOI":"10.1038\/s42003-018-0186-4","type":"journal-article","created":{"date-parts":[[2018,10,29]],"date-time":"2018-10-29T15:40:46Z","timestamp":1540827646000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Substrate specificity of plant nitrilase complexes is affected by their helical twist"],"prefix":"10.1038","volume":"1","author":[{"given":"Jeremy D.","family":"Woodward","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inga","family":"Trompetter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6701-2394","authenticated-orcid":false,"given":"B. Trevor","family":"Sewell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6347-3337","authenticated-orcid":false,"given":"Markus","family":"Piotrowski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,11,2]]},"reference":[{"key":"186_CR1","doi-asserted-by":"publisher","first-page":"1344","DOI":"10.1002\/pro.5560030821","volume":"3","author":"P Bork","year":"1994","unstructured":"Bork, P. & Koonin, E. V. A new family of carbon-nitrogen hydrolases. Protein Sci. 3, 1344\u20131346 (1994).","journal-title":"Protein Sci."},{"key":"186_CR2","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2001-2-1-reviews0001","volume":"2","author":"HC Pace","year":"2001","unstructured":"Pace, H. C. & Brenner, C. The nitrilase superfamily: classification, structure and function. Genome Biol. 2, REVIEWS0001 (2001).","journal-title":"Genome Biol."},{"key":"186_CR3","doi-asserted-by":"publisher","first-page":"1161","DOI":"10.1046\/j.1365-2672.2003.02123.x","volume":"95","author":"C O\u2019Reilly","year":"2003","unstructured":"O\u2019Reilly, C. & Turner, P. D. The nitrilase family of CN hydrolysing enzymes - a comparative study. J. Appl. Microbiol. 95, 1161\u20131174 (2003).","journal-title":"J. Appl. Microbiol."},{"key":"186_CR4","doi-asserted-by":"publisher","first-page":"2655","DOI":"10.1016\/j.phytochem.2008.08.020","volume":"69","author":"M Piotrowski","year":"2008","unstructured":"Piotrowski, M. Primary or secondary? Versatile nitrilases in plant metabolism. Phytochemistry 69, 2655\u20132667 (2008).","journal-title":"Phytochemistry"},{"key":"186_CR5","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1111\/j.1365-2672.2008.03941.x","volume":"106","author":"RN Thuku","year":"2009","unstructured":"Thuku, R. N., Brady, D., Benedik, M. J. & Sewell, B. T. Microbial nitrilases: versatile, spiral forming, industrial enzymes. J. Appl. Microbiol. 106, 703\u2013727 (2009).","journal-title":"J. Appl. Microbiol."},{"key":"186_CR6","doi-asserted-by":"publisher","first-page":"1413","DOI":"10.1016\/j.str.2003.10.005","volume":"11","author":"BT Sewell","year":"2003","unstructured":"Sewell, B. T., Berman, M. N., Meyers, P. R., Jandhyala, D. & Benedik, M. J. The cyanide degrading nitrilase from Pseudomonas stutzeri AK61 is a two-fold symmetric, 14-subunit spiral. Structure 11, 1413\u20131422 (2003).","journal-title":"Structure"},{"key":"186_CR7","doi-asserted-by":"publisher","first-page":"2099","DOI":"10.1111\/j.1742-4658.2007.05752.x","volume":"274","author":"RN Thuku","year":"2007","unstructured":"Thuku, R. N., Weber, B. W., Varsani, A. & Sewell, B. T. Post-translational cleavage of recombinantly expressed nitrilase from Rhodococcus rhodochrous J1 yields a stable, active helical form. FEBS J. 274, 2099\u20132108 (2007).","journal-title":"FEBS J."},{"key":"186_CR8","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1016\/j.molcatb.2007.09.006","volume":"50","author":"V Vejvoda","year":"2008","unstructured":"Vejvoda, V. et al. Purification and characterization of a nitrilase from Fusarium solani O1. J. Mol. Catal. B-Enzym 50, 99\u2013106 (2008).","journal-title":"J. Mol. Catal. B-Enzym"},{"key":"186_CR9","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.jsb.2007.09.019","volume":"161","author":"JD Woodward","year":"2008","unstructured":"Woodward, J. D. et al. Helical structure of unidirectionally shadowed metal replicas of cyanide hydratase from Gloeocercospora sorghi. J. Struct. Biol. 161, 111\u2013119 (2008).","journal-title":"J. Struct. Biol."},{"key":"186_CR10","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1007\/s00253-008-1735-4","volume":"82","author":"KC Dent","year":"2009","unstructured":"Dent, K. C., Weber, B. W., Benedik, M. J. & Sewell, B. T. The cyanide hydratase from Neurospora crassa forms a helix which has a dimeric repeat. Appl. Microbiol. Biotechnol. 82, 271\u2013278 (2009).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"186_CR11","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1007\/s00253-010-2734-9","volume":"88","author":"DS Williamson","year":"2010","unstructured":"Williamson, D. S. et al. Structural and biochemical characterization of a nitrilase from the thermophilic bacterium, Geobacillus pallidus RAPc8. Appl. Microbiol. Biotechnol. 88, 143\u2013153 (2010).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"186_CR12","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1007\/s10529-010-0421-7","volume":"33","author":"O Kaplan","year":"2011","unstructured":"Kaplan, O. et al. Genome mining for the discovery of new nitrilases in filamentous fungi. Biotechnol. Lett. 33, 309\u2013312 (2011).","journal-title":"Biotechnol. Lett."},{"key":"186_CR13","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1111\/nph.12185","volume":"198","author":"A Doskocilova","year":"2013","unstructured":"Doskocilova, A. et al. NITRILASE1 regulates the exit from proliferation, genome stability and plant development. New Phytol. 198, 685\u2013698 (2013).","journal-title":"New Phytol."},{"key":"186_CR14","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1046\/j.1432-1327.2000.00983.x","volume":"267","author":"T Nagasawa","year":"2000","unstructured":"Nagasawa, T. et al. Nitrilase of Rhodococcus rhodochrous J1. Conversion into the active form by subunit association. Eur. J. Biochem. 267, 138\u2013144 (2000).","journal-title":"Eur. J. Biochem."},{"key":"186_CR15","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1007\/s00253-005-1903-8","volume":"68","author":"DM Jandhyala","year":"2005","unstructured":"Jandhyala, D. M., Willson, R. C., Sewell, B. T. & Benedik, M. J. Comparison of cyanide-degrading nitrilases. Appl. Microbiol. Biotechnol. 68, 327\u2013335 (2005).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"186_CR16","doi-asserted-by":"publisher","first-page":"18848","DOI":"10.1073\/pnas.0709315104","volume":"104","author":"R Jenrich","year":"2007","unstructured":"Jenrich, R. et al. Evolution of heteromeric nitrilase complexes in Poaceae with new functions in nitrile metabolism. Proc. Natl Acad. Sci. USA 104, 18848\u201318853 (2007).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"186_CR17","doi-asserted-by":"publisher","first-page":"1279","DOI":"10.1002\/cbdv.200690131","volume":"3","author":"R Singh","year":"2006","unstructured":"Singh, R., Sharma, R., Tewari, N. & Rawat, D. S. Nitrilase and its application as a \u2018green\u2019 catalyst. Chem. Biodivers. 3, 1279\u20131287 (2006).","journal-title":"Chem. Biodivers."},{"key":"186_CR18","doi-asserted-by":"publisher","first-page":"142","DOI":"10.1186\/1475-2859-11-142","volume":"11","author":"JS Gong","year":"2012","unstructured":"Gong, J. S. et al. Nitrilases in nitrile biocatalysis: recent progress and forthcoming research. Microb. Cell. Fact. 11, 142 (2012).","journal-title":"Microb. Cell. Fact."},{"key":"186_CR19","doi-asserted-by":"publisher","first-page":"5592","DOI":"10.1128\/AEM.00301-09","volume":"75","author":"C Kiziak","year":"2009","unstructured":"Kiziak, C. & Stolz, A. Identification of amino acid residues responsible for the enantioselectivity and amide formation capacity of the Arylacetonitrilase from Pseudomonas fluorescens EBC191. Appl. Environ. Microbiol. 75, 5592\u20135599 (2009).","journal-title":"Appl. Environ. Microbiol."},{"key":"186_CR20","first-page":"384","volume":"3","author":"S Kaushik","year":"2012","unstructured":"Kaushik, S., Mohan, U. & Banerjee, U. Exploring residues crucial for nitrilase function by site directed mutagenesis to gain better insight into sequence-function relationships. Int. J. Biochem. Mol. Biol. 3, 384\u2013391 (2012).","journal-title":"Int. J. Biochem. Mol. Biol."},{"key":"186_CR21","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.jsb.2014.10.003","volume":"188","author":"L Zhang","year":"2014","unstructured":"Zhang, L. et al. Structural insights into enzymatic activity and substrate specificity determination by a single amino acid in nitrilase from Syechocystis sp. PCC6803. J. Struct. Biol. 188, 93\u2013101 (2014).","journal-title":"J. Struct. Biol."},{"key":"186_CR22","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1007\/s00449-014-1258-6","volume":"38","author":"H Li","year":"2015","unstructured":"Li, H. et al. Improving the catalytic potential and substrate tolerance of Gibberella intermedia nitrilase by whole-cell immobilization. Bioprocess Biosyst. Eng. 38, 189\u2013197 (2015).","journal-title":"Bioprocess Biosyst. Eng."},{"key":"186_CR23","doi-asserted-by":"publisher","first-page":"69","DOI":"10.3109\/07388551.2015.1120704","volume":"37","author":"JS Gong","year":"2017","unstructured":"Gong, J. S. et al. Nitrile-converting enzymes as a tool to improve biocatalysis in organic synthesis: recent insights and promises. Crit. Rev. Biotechnol. 37, 69\u201381 (2017).","journal-title":"Crit. Rev. Biotechnol."},{"key":"186_CR24","doi-asserted-by":"publisher","first-page":"2616","DOI":"10.1074\/jbc.M007890200","volume":"276","author":"M Piotrowski","year":"2001","unstructured":"Piotrowski, M., Sch\u00f6nfelder, S. & Weiler, E. W. The Arabidopsis thaliana isogene NIT4 and its orthologs in tobacco encode \u00df-cyano-L-alanine hydratase\/nitrilase. J. Biol. Chem. 276, 2616\u20132621 (2001).","journal-title":"J. Biol. Chem."},{"key":"186_CR25","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1007\/s11103-005-6217-9","volume":"61","author":"M Piotrowski","year":"2006","unstructured":"Piotrowski, M. & Volmer, J. J. Cyanide metabolism in higher plants: cyanoalanine hydratase is a NIT4 homolog. Plant Mol. Biol. 61, 111\u2013122 (2006).","journal-title":"Plant Mol. Biol."},{"key":"186_CR26","doi-asserted-by":"publisher","first-page":"1680","DOI":"10.1016\/j.phytochem.2009.07.028","volume":"70","author":"T Janowitz","year":"2009","unstructured":"Janowitz, T., Trompetter, I. & Piotrowski, M. Evolution of nitrilases in glucosinolate-containing plants. Phytochemistry 70, 1680\u20131686 (2009).","journal-title":"Phytochemistry"},{"key":"186_CR27","doi-asserted-by":"publisher","first-page":"508","DOI":"10.1007\/s004250000420","volume":"212","author":"S Vorwerk","year":"2001","unstructured":"Vorwerk, S. et al. Enzymatic characterization of the recombinant Arabidopsis thaliana nitrilase subfamily encoded by the NIT2\/NIT1\/NIT3-gene cluster. Planta 212, 508\u2013516 (2001).","journal-title":"Planta"},{"key":"186_CR28","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1111\/j.1432-1033.1992.tb16795.x","volume":"205","author":"D Bartling","year":"1992","unstructured":"Bartling, D., Seedorf, M., Mithofer, A. & Weiler, E. W. Cloning and expression of an Arabidopsis nitrilase which can convert indole-3-acetonitrile to the plant hormone, indole-3-acetic acid. Eur. J. Biochem. 205, 417\u2013424 (1992).","journal-title":"Eur. J. Biochem."},{"key":"186_CR29","doi-asserted-by":"publisher","first-page":"6649","DOI":"10.1073\/pnas.91.14.6649","volume":"91","author":"B Bartel","year":"1994","unstructured":"Bartel, B. & Fink, G. R. Differential regulation of an auxin-producing nitrilase gene family in Arabidopsis thaliana. Proc. Natl Acad. Sci. USA 91, 6649\u20136653 (1994).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"186_CR30","doi-asserted-by":"publisher","unstructured":"Hashimoto, H., et al Crystal structure of C171A\/V236A mutant of N-carbamyl-D-amino acid amidohydrolase. RCSB Protein Data Bank. https:\/\/doi.org\/10.2210\/pdb1uf5\/pdb (2003).","DOI":"10.2210\/pdb1uf5\/pdb"},{"key":"186_CR31","doi-asserted-by":"publisher","first-page":"19598","DOI":"10.1074\/jbc.M701039200","volume":"282","author":"J Andrade","year":"2007","unstructured":"Andrade, J., Karmali, A., Carrondo, M. A. & Frazao, C. Structure of amidase from Pseudomonas aeruginosa showing a trapped acyl transfer reaction intermediate state. J. Biol. Chem. 282, 19598\u201319605 (2007).","journal-title":"J. Biol. Chem."},{"key":"186_CR32","doi-asserted-by":"publisher","unstructured":"Kimani, S. W., Hunter, R., Vlok, M., Watermeyer, J. & Sewell, B. T. Covalent modifications of the active site cysteine occur as a result of mutating the glutamate of the catalytic triad in the amidase from Nesterenkonia sp. RCSB Protein Data Bank. https:\/\/doi.org\/10.2210\/pdb4izs\/pdb (2013).","DOI":"10.2210\/pdb4izs\/pdb"},{"key":"186_CR33","doi-asserted-by":"publisher","first-page":"4794","DOI":"10.1128\/AEM.69.8.4794-4805.2003","volume":"69","author":"D Jandhyala","year":"2003","unstructured":"Jandhyala, D. et al. CynD, the cyanide dihydratase from Bacillus pumilus: gene cloning and structural studies. Appl. Environ. Microbiol. 69, 4794\u20134805 (2003).","journal-title":"Appl. Environ. Microbiol."},{"key":"186_CR34","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1007\/s00253-005-0255-8","volume":"72","author":"A Banerjee","year":"2006","unstructured":"Banerjee, A., Kaul, P. & Banerjee, U. C. Enhancing the catalytic potential of nitrilase from Pseudomonas putida for stereoselective nitrile hydrolysis. Appl. Microbiol. Biotechnol. 72, 77\u201387 (2006).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"186_CR35","doi-asserted-by":"publisher","first-page":"1544","DOI":"10.1016\/j.jmb.2008.02.011","volume":"377","author":"S Lundgren","year":"2008","unstructured":"Lundgren, S., Lohkamp, B., Andersen, B., Piskur, J. & Dobritzsch, D. The crystal structure of \u00df-alanine synthase from Drosophila melanogaster reveals a homooctameric helical turn-like assembly. J. Mol. Biol. 377, 1544\u20131559 (2008).","journal-title":"J. Mol. Biol."},{"key":"186_CR36","doi-asserted-by":"publisher","first-page":"1761","DOI":"10.1093\/bioinformatics\/btp302","volume":"25","author":"A Lobley","year":"2009","unstructured":"Lobley, A., Sadowski, M. I. & Jones, D. T. pGenTHREADER and pDomTHREADER: new methods for improved protein fold recognition and superfamily discrimination. Bioinformatics 25, 1761\u20131767 (2009).","journal-title":"Bioinformatics"},{"key":"186_CR37","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1006\/jmbi.2001.4762","volume":"310","author":"JY Shi","year":"2001","unstructured":"Shi, J. Y., Blundell, T. L. & Mizuguchi, K. FUGUE: Sequence-structure homology recognition using environment-specific substitution tables and structure-dependent gap penalties. J. Mol. Biol. 310, 243\u2013257 (2001).","journal-title":"J. Mol. Biol."},{"key":"186_CR38","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1002\/0471250953.bi0506s47","volume":"47","author":"B Webb","year":"2014","unstructured":"Webb, B. & Sali, A. Comparative protein structure modeling using MODELLER. Curr. Protoc. Bioinforma. 47, 1\u201332 (2014).","journal-title":"Curr. Protoc. Bioinforma."},{"key":"186_CR39","doi-asserted-by":"publisher","first-page":"1605","DOI":"10.1002\/jcc.20084","volume":"25","author":"EF Pettersen","year":"2004","unstructured":"Pettersen, E. F. et al. UCSF Chimera - a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605\u20131612 (2004).","journal-title":"J. Comput. Chem."},{"key":"186_CR40","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1023\/A:1008848517221","volume":"12","author":"T Wurch","year":"1998","unstructured":"Wurch, T., Lestienne, F. & Pauwels, P. J. A modified overlap extension PCR method to create chimeric genes in the absence of restriction enzymes. Biotechnol. Tech. 12, 653\u2013657 (1998).","journal-title":"Biotechnol. Tech."},{"key":"186_CR41","doi-asserted-by":"publisher","first-page":"e115","DOI":"10.1093\/nar\/gnh110","volume":"32","author":"L Zheng","year":"2004","unstructured":"Zheng, L., Baumann, U. & Reymond, J. L. An efficient one-step site-directed and site-saturation mutagenesis protocol. Nucleic Acids Res. 32, e115 (2004).","journal-title":"Nucleic Acids Res."},{"key":"186_CR42","first-page":"E3227","volume":"58","author":"DS Booth","year":"2011","unstructured":"Booth, D. S., Avila-Sakar, A. & Cheng, Y. F. Visualizing proteins and macromolecular complexes by negative stain EM: from grid preparation to image acquisition. J. Vis. Exp. 58, E3227 (2011).","journal-title":"J. Vis. Exp."},{"key":"186_CR43","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/S0304-3991(00)00062-0","volume":"85","author":"EH Egelman","year":"2000","unstructured":"Egelman, E. H. A robust algorithm for the reconstruction of helical filaments using single-particle methods. Ultramicroscopy 85, 225\u2013234 (2000).","journal-title":"Ultramicroscopy"},{"key":"186_CR44","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1006\/jsbi.1996.0030","volume":"116","author":"J Frank","year":"1996","unstructured":"Frank, J. et al. SPIDER and WEB: processing and visualization of images in 3D electron microscopy and related fields. J. Struct. Biol. 116, 190\u2013199 (1996).","journal-title":"J. Struct. Biol."},{"key":"186_CR45","doi-asserted-by":"publisher","first-page":"791","DOI":"10.1107\/S090744490000679X","volume":"56","author":"ML Quillin","year":"2000","unstructured":"Quillin, M. L. & Matthews, B. W. Accurate calculation of the density of proteins. Acta Crystallogr D 56, 791\u2013794 (2000).","journal-title":"Acta Crystallogr D"},{"key":"186_CR46","doi-asserted-by":"publisher","first-page":"28514","DOI":"10.1074\/jbc.M113.503284","volume":"288","author":"BW Weber","year":"2013","unstructured":"Weber, B. W. et al. The mechanism of the amidases: mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning. J. Biol. Chem. 288, 28514\u201328523 (2013).","journal-title":"J. Biol. Chem."}],"container-title":["Communications Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s42003-018-0186-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-018-0186-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-018-0186-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T12:57:49Z","timestamp":1671627469000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s42003-018-0186-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,2]]},"references-count":46,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["186"],"URL":"https:\/\/doi.org\/10.1038\/s42003-018-0186-4","relation":{},"ISSN":["2399-3642"],"issn-type":[{"value":"2399-3642","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,2]]},"assertion":[{"value":"9 March 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 October 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 November 2018","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"186"}}