{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T15:13:31Z","timestamp":1761059611444,"version":"build-2065373602"},"reference-count":29,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2013,10,15]],"date-time":"2013-10-15T00:00:00Z","timestamp":1381795200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Calcineurin (or PP2B) has been reported to be involved in an array of physiological process in insects, and the calcineurin subunit A (CNA) plays a central role  in calcineurin activity. We cloned the CNA gene from Plutella xylostella (PxCNA). This gene contains an ORF of 1488 bp that encodes a 495 amino acid protein, showing 98%, and 80% identities to the CNA of Bombyx mori, and humans respectively. The full-length of PxCNA and its catalytic domain (CNA1\u2013341, defined as PxCN\u03b1) were both expressed in Escherichia coli. Purified recombinant PxCNA displayed no phosphatase activity, whereas recombinant PxCN\u03b1 showed high phosphatase activity with a Km of 4.6 mM and a kcat  of 0.66 S\u22121 against pNPP. It could be activated at different degrees by Mn2+, Ni2+, Mg2+, and Ca2+. The optimum reaction pH was about 7.5 and the optimum reaction temperature was around 45 \u00b0C. An in vitro inhibition assay showed that okadaic acid (OA) and cantharidin (CTD) competitively inhibited recombinant PxCN\u03b1 activity with the IC50 values of 8.95 \u03bcM and 77.64 \u03bcM, respectively. However, unlike previous reports, pyrethroid insecticides were unable to inhibit recombinant PxCN\u03b1, indicating that the  P. xylostella calcineurin appears not to be sensitive to class II pyrethroid insecticides.<\/jats:p>","DOI":"10.3390\/ijms141020692","type":"journal-article","created":{"date-parts":[[2013,10,15]],"date-time":"2013-10-15T12:47:44Z","timestamp":1381841264000},"page":"20692-20703","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Molecular Cloning and Characterization of the Calcineurin Subunit A from Plutella xylostella"],"prefix":"10.3390","volume":"14","author":[{"given":"Xi'en","family":"Chen","sequence":"first","affiliation":[{"name":"Key Laboratory of Plant Protection Resources & Pest Management of Ministry of Education, Northwest A&F University, Yangling 712100, Shaanxi, China"}]},{"given":"Yalin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources & Pest Management of Ministry of Education, Northwest A&F University, Yangling 712100, Shaanxi, China"}]}],"member":"1968","published-online":{"date-parts":[[2013,10,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1152\/physrev.2000.80.4.1483","article-title":"Calcineurin: Form and function","volume":"80","author":"Rusnak","year":"2000","journal-title":"Physiol. Rev"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/S0070-2137(01)80011-X","article-title":"Calcineurin: From structure to function","volume":"36","author":"Aramburu","year":"2001","journal-title":"Curr. Top. Cell. Regul"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/0092-8674(95)90439-5","article-title":"X-ray structure of calcineurin inhibited by the immunophilin-immunosuppressant FKBP12-FK506 complex","volume":"82","author":"Grifith","year":"1995","journal-title":"Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1868","DOI":"10.1021\/bi00430a066","article-title":"Functional domain structure of calcineurin A: Mapping by limited proteolysis","volume":"28","author":"Hubbard","year":"1989","journal-title":"Biochemistry"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/0014-5793(94)80398-6","article-title":"Identification of a cDNA encoding a Drosophila calcium\/calmodulin regulated protein phosphatase, which has its most abundant expression in the early embryo","volume":"339","author":"Brown","year":"1994","journal-title":"FEBS Lett"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/S0965-1748(01)00125-4","article-title":"cDNA cloning of calcineurin heterosubunits from the pheromone gland of the silk moth Bombyx mori","volume":"32","author":"Yoshiga","year":"2002","journal-title":"Insect Biochem. Mol. Biol"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/S0305-0491(99)00096-6","article-title":"Involvement of calcineurin in the signal transduction of PBAN in the silkworm, Bombyx mori (Lepidoptera)","volume":"124","author":"Yokoyama","year":"1999","journal-title":"Comp. Biochem. Phys. B"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/S0168-0102(96)01132-7","article-title":"An inhibitory role of calcineurin in endocytosis of synaptic vesicles at nerve terminals of Drosophila larvae","volume":"27","author":"Kuromi","year":"1997","journal-title":"Neurosci. Res"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1016\/j.ydbio.2010.06.011","article-title":"Calcineurin and its regulation by Sra\/RCAN is required for completion of meiosis in Drosophila","volume":"344","author":"Takeo","year":"2010","journal-title":"Dev. Biol"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1073\/pnas.0337662100","article-title":"Requirement of the calcineurin subunit gene canB2 for indirect flight muscle formation in Drosophila","volume":"100","author":"Gajewski","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"12759","DOI":"10.1523\/JNEUROSCI.1337-11.2011","article-title":"Calcineurin and its regulator sra\/DSCR1 are essential for sleep in Drosophila","volume":"31","author":"Nakai","year":"2011","journal-title":"J. Neurosci"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13137","DOI":"10.1523\/JNEUROSCI.5860-10.2011","article-title":"Pan-neuronal knockdown of calcineurin reduces sleep in the fruit fly Drosophila melanogaster","volume":"31","author":"Tomita","year":"2011","journal-title":"J. Neurosci"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"C1047","DOI":"10.1152\/ajpcell.00328.2009","article-title":"Calcineurin activity augments cAMP\/PKA-dependent activation of V-ATPase in blowfly salivary glands","volume":"298","author":"Voss","year":"2010","journal-title":"Am. J. Physiol. Cell Physiol"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"22542","DOI":"10.1016\/S0021-9258(18)41706-1","article-title":"Molecular cloning and characterization of the genes encoding the two subunits of Drosophila melanogaster calcineurin","volume":"267","author":"Guerini","year":"1992","journal-title":"J. Biol. Chem"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.biochi.2004.10.009","article-title":"Non-catalytic domains of subunit A negatively regulate the activity of calcineurin","volume":"87","author":"Liu","year":"2005","journal-title":"Biochemistry"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1429","DOI":"10.1515\/BC.2003.158","article-title":"The catalytically active domain in the A subunit of calcineurin","volume":"384","author":"Xiang","year":"2003","journal-title":"Biol. Chem"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1023\/B:BIOM.0000018373.85342.36","article-title":"Effect of metal ions on the activity of the catalytic domain of calcineurin","volume":"17","author":"Liu","year":"2004","journal-title":"Biometals"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6134","DOI":"10.1016\/S0021-9258(20)82115-2","article-title":"Regulation of calcineurin by metal ions. Mechanism of activation by Ni2+ and an enhanced response to Ca2+\/calmodulin","volume":"259","author":"Pallen","year":"1984","journal-title":"J. Biol. Chem"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"16115","DOI":"10.1016\/S0021-9258(18)66685-2","article-title":"Stoichiometry and dynamic interaction of metal ion activators with calcineurin phosphatase","volume":"261","author":"Pallen","year":"1986","journal-title":"J. Biol. Chem"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3386","DOI":"10.1021\/bi981748l","article-title":"Comparison of the reaction progress of calcineurin with Mn2+ and Mg2+","volume":"38","author":"Martin","year":"1999","journal-title":"Biochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1016\/0006-2952(92)90710-Z","article-title":"Specific inhibition of calcineurin by type II synthetic pyrethroid insecticides","volume":"43","author":"Enan","year":"1992","journal-title":"Biochem. Pharmacol"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/1471-2121-2-6","article-title":"Serine\/threonine protein phosphatase 5 (PP5) participates in the regulation of glucocorticoid receptor nucleocytoplasmic shuttling","volume":"2","author":"Dean","year":"2001","journal-title":"BMC Cell Biol"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.2174\/0929867023368836","article-title":"Regulators of serine\/threonine protein phosphatases at the dawn of a clinical era?","volume":"9","author":"Honkanen","year":"2002","journal-title":"Curr. Med. Chem"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"44078","DOI":"10.1074\/jbc.M107656200","article-title":"Crystal structure of the tumor-promoter okadaic acid bound to protein phosphatase-1","volume":"276","author":"Maynes","year":"2001","journal-title":"J. Biol. Chem"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.cell.2006.09.025","article-title":"Structure of protein phosphatase 2A core enzyme bound to tumor-inducing toxins","volume":"127","author":"Xing","year":"2006","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4838","DOI":"10.1021\/jm900610k","article-title":"Structural basis of serine\/threonine phosphatase inhibition by the archetypal small molecules cantharidin and norcantharidin","volume":"52","author":"Bertini","year":"2009","journal-title":"J. Med. Chem"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1111\/j.1399-3054.2011.01494.x","article-title":"Transcriptional responses to cantharidin, a protein phosphatase inhibitor, in Arabidopsis thaliana reveal the involvement of multiple signal transduction pathways","volume":"143","author":"Bajsa","year":"2011","journal-title":"Physiol. Plant"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1080\/15216549700201171","article-title":"Expression and reconstitution of calcineurin A and B subunits","volume":"41","author":"Wei","year":"1997","journal-title":"IUBMB Life"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/14\/10\/20692\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:49:52Z","timestamp":1760219392000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/14\/10\/20692"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,10,15]]},"references-count":29,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2013,10]]}},"alternative-id":["ijms141020692"],"URL":"https:\/\/doi.org\/10.3390\/ijms141020692","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2013,10,15]]}}}