{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T01:28:06Z","timestamp":1774747686077,"version":"3.50.1"},"reference-count":51,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2018,10,29]],"date-time":"2018-10-29T00:00:00Z","timestamp":1540771200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001023","name":"Australian Institute of Nuclear Science and Engineering","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001023","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Neurochemistry"],"published-print":{"date-parts":[[2018,11]]},"abstract":"Abstract<\/jats:title>The Parkinson's disease (PD<\/jats:styled-content>)\u2010causative leucine\u2010rich repeat kinase 2 (LRRK<\/jats:styled-content>2) belongs to the Roco family of G\u2010proteins comprising a Ras\u2010of\u2010complex (Roc) domain followed by a C\u2010terminal of Roc (COR<\/jats:styled-content>) domain in tandem (called Roc\u2010COR<\/jats:styled-content> domain). Two prokaryotic Roc\u2010COR<\/jats:styled-content> domains have been characterized as \u2018G proteins activated by guanine nucleotide\u2010dependent dimerization\u2019 (GAD<\/jats:styled-content>s), which require dimerization for activation of their GTP<\/jats:styled-content>ase activity and bind guanine nucleotides with relatively low affinities. Additionally, LRRK<\/jats:styled-content>2 Roc domain in isolation binds guanine nucleotides with relatively low affinities. As such, LRRK<\/jats:styled-content>2 GTP<\/jats:styled-content>ase domain was predicted to be a GAD<\/jats:styled-content>. Herein, we describe the design and high\u2010level expression of human LRRK<\/jats:styled-content>2 Roc\u2010COR<\/jats:styled-content> domain (LRRK<\/jats:styled-content>2 Roc\u2010COR<\/jats:styled-content>). Biochemical analyses of LRRK<\/jats:styled-content>2 Roc\u2010COR<\/jats:styled-content> reveal that it forms homodimers, with the C\u2010terminal portion of COR<\/jats:styled-content> mediating its dimerization. Furthermore, it co\u2010purifies and binds Mg2+<\/jats:sup>GTP<\/jats:styled-content>\/GDP<\/jats:styled-content> at 1\u00a0:\u00a01 stoichiometry, and it hydrolyzes GTP<\/jats:styled-content> with Km<\/jats:sub>\u00a0and\u00a0k<\/jats:italic>cat\u00a0<\/jats:sub>of 22 nM<\/jats:styled-content> and 4.70\u00a0\u00d7\u00a010\u22124<\/jats:sup>\u00a0min\u22121<\/jats:sup>,\u00a0<\/jats:sub>respectively. Thus, even though LRRK<\/jats:styled-content>2 Roc\u2010COR<\/jats:styled-content> forms GAD<\/jats:styled-content>\u2010like homodimers, it exhibits conventional Ras\u2010like GTP<\/jats:styled-content>ase properties, with high\u2010affinity binding of Mg2+<\/jats:sup>\u2010GTP<\/jats:styled-content>\/GDP<\/jats:styled-content> and low intrinsic catalytic activity. The PD<\/jats:styled-content>\u2010causative Y1699C mutation mapped to the COR<\/jats:styled-content> domain was previously reported to reduce the GTP<\/jats:styled-content>ase activity of full\u2010length LRRK<\/jats:styled-content>2. In contrast, this mutation induces no change in the GTP<\/jats:styled-content>ase activity, and only slight perturbations in the secondary structure contents of LRRK<\/jats:styled-content>2 Roc\u2010COR<\/jats:styled-content>. As this mutation does not directly affect the GTP<\/jats:styled-content>ase activity of the isolated Roc\u2010COR<\/jats:styled-content> tandem, it is possible that the effects of this mutation on full\u2010length LRRK<\/jats:styled-content>2 occur via other functional domains.<\/jats:p>Open Practices<\/jats:title><\/jats:p>Open Science: This manuscript was awarded with the Open Materials Badge.<\/jats:p>For more information see: https:\/\/cos.io\/our-services\/open-science-badges\/<\/jats:ext-link><\/jats:p><\/jats:sec><\/jats:sec>\nimage<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text>\n<\/jats:p><\/jats:sec>","DOI":"10.1111\/jnc.14566","type":"journal-article","created":{"date-parts":[[2018,8,9]],"date-time":"2018-08-09T06:49:57Z","timestamp":1533797397000},"page":"409-428","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["The Roc\u2010COR tandem domain of leucine\u2010rich repeat kinase 2 forms dimers and exhibits conventional Ras\u2010like GTPase properties"],"prefix":"10.1111","volume":"147","author":[{"given":"Ryan D.","family":"Mills","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lung\u2010Yu","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"},{"name":"Cell Signaling Research Laboratories University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daisy Sio\u2010Seng","family":"Lio","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"},{"name":"Cell Signaling Research Laboratories University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yee\u2010Foong","family":"Mok","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Terrence D.","family":"Mulhern","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George","family":"Cao","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Griffin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vijaya B.","family":"Kenche","sequence":"additional","affiliation":[{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"},{"name":"Florey Neuroscience Institute University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janetta G.","family":"Culvenor","sequence":"additional","affiliation":[{"name":"Department of Pathology University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4965-7148","authenticated-orcid":false,"given":"Heung\u2010Chin","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Melbourne Parkville Victoria Australia"},{"name":"Bio21 Institute of Molecular Science and Biotechnology University of Melbourne Parkville Victoria Australia"},{"name":"Cell Signaling Research Laboratories University of Melbourne Parkville Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2018,10,29]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2008.06789.x"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/dds522"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2003.08.008"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2010.05.009"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0043472"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mad.2010.01.009"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2010.07105.x"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0709098105"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-01103-4"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2011.07537.x"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.parkreldis.2008.06.009"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(81)81118-0"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm2689"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2008.150"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1523708113"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2007.07.007"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.192453199"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddv314"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20111215"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi061960m"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi400596m"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2009.06358.x"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2007.04.006"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.2007.04743.x"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbadis.2009.09.010"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0017153"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1323285111"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi901851y"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi201173d"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.15-277095"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.08-111310"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tins.2006.03.006"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2012.08.022"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1500851112"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20120088"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.22515"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20100483"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20160240"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2004.10.023"},{"key":"e_1_2_8_41_1","doi-asserted-by":"crossref","first-page":"3906","DOI":"10.1016\/S0021-9258(18)89207-9","article-title":"Viral Harvey ras p21 expressed in Escherichia coli purifies as a binary one\u2010to\u2010one complex with GDP","volume":"260","author":"Poe M.","year":"1985","journal-title":"J. Biol. Chem."},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1042\/BSR20150128"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-09126-z"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.025437"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(05)07002-3"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0023207"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20141095"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20160224"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-062708-134043"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1000902"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.4566-11.2012"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M001027200"}],"container-title":["Journal of Neurochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fjnc.14566","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jnc.14566","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T00:19:01Z","timestamp":1694737141000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/jnc.14566"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,29]]},"references-count":51,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2018,11]]}},"alternative-id":["10.1111\/jnc.14566"],"URL":"https:\/\/doi.org\/10.1111\/jnc.14566","archive":["Portico"],"relation":{},"ISSN":["0022-3042","1471-4159"],"issn-type":[{"value":"0022-3042","type":"print"},{"value":"1471-4159","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10,29]]},"assertion":[{"value":"2017-12-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-08-03","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-10-29","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}