{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,27]],"date-time":"2026-06-27T11:55:51Z","timestamp":1782561351726,"version":"3.54.5"},"reference-count":39,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2018,3,31]],"date-time":"2018-03-31T00:00:00Z","timestamp":1522454400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the China Natural Science Foundation.","award":["81173035"],"award-info":[{"award-number":["81173035"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>\u03b1-Conotoxins (\u03b1-CTxs) are small peptides composed of 11 to 20 amino acid residues with two disulfide bridges. Most of them potently and selectively target nicotinic acetylcholine receptor (nAChR) subtypes, and a few were found to inhibit the GABAB receptor (GABABR)-coupled N-type calcium channels (Cav2.2). However, in all of \u03b1-CTxs targeting both receptors, the disulfide connectivity arrangement \u201cC1-C3, C2-C4\u201d is present. In this work, a novel \u03b14\/7-CTx named Lt1.3 (GCCSHPACSGNNPYFC-NH2) was cloned from the venom ducts of Conus litteratus (C. litteratus) in the South China Sea. Lt1.3 was then chemically synthesized and two isomers with disulfide bridges \u201cC1-C3, C2-C4\u201d and \u201cC1-C4, C2-C3\u201d were found and functionally characterized. Electrophysiological experiments showed that Lt1.3 containing the common disulfide bridges \u201cC1-C3, C2-C4\u201d potently and selectively inhibited \u03b13\u03b22 nAChRs and not GABABR-coupled Cav2.2. Surprisingly, but the isomer with the disulfide bridges \u201cC1-C4, C2-C3\u201d showed exactly the opposite inhibitory activity, inhibiting only GABABR-coupled Cav2.2 and not \u03b13\u03b22 nAChRs. These findings expand the knowledge of the targets and selectivity of \u03b1-CTxs and provide a new structural motif to inhibit the GABABR-coupled Cav2.2.<\/jats:p>","DOI":"10.3390\/md16040112","type":"journal-article","created":{"date-parts":[[2018,4,2]],"date-time":"2018-04-02T12:32:20Z","timestamp":1522672340000},"page":"112","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Cloning, Synthesis and Functional Characterization of a Novel \u03b1-Conotoxin Lt1.3"],"prefix":"10.3390","volume":"16","author":[{"given":"Jinqin","family":"Chen","sequence":"first","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"},{"name":"Institute of Physical Science and Information Technology, Anhui University, Hefei 236041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Liang","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huying","family":"Ning","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengtao","family":"Cai","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuguo","family":"Liu","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Longxiao","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangyi","family":"Zhou","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2062-8332","authenticated-orcid":false,"given":"Qiuyun","family":"Dai","sequence":"additional","affiliation":[{"name":"Beijing Institute of Biotechnology, Beijing 100071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6058","DOI":"10.3390\/md12126058","article-title":"Conotoxin gene superfamilies","volume":"12","author":"Robinson","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"D325","DOI":"10.1093\/nar\/gkr886","article-title":"ConoServer: Updated content, knowledge, and discovery tools in the conopeptide database","volume":"40","author":"Kaas","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5386","DOI":"10.3390\/toxins7124890","article-title":"Conotoxins that could provide analgesia through voltage gated sodium channel inhibition","volume":"7","author":"Munasinghe","year":"2015","journal-title":"Toxins"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"5815","DOI":"10.1021\/cr400401e","article-title":"Discovery, synthesis, and structure-activity relationships of conotoxins","volume":"114","author":"Akondi","year":"2014","journal-title":"Chem. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Ram\u00edrez, D., Gonzalez, W., Fissore, R.A., and Carvacho, I. (2017). Conotoxins as tools to understand the physiological function of voltage-gated calcium (Ca(V)) channels. Mar. Drugs, 15.","DOI":"10.3390\/md15100313"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2970","DOI":"10.3390\/md12052970","article-title":"Conotoxins targeting nicotinic acetylcholine receptors: An overview","volume":"12","author":"Lebbe","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"17837","DOI":"10.1074\/jbc.M610813200","article-title":"Chi-Conotoxin and tricyclic antidepressant interactions at the norepinephrine transporter define a new transporter model","volume":"282","author":"Paczkowski","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"633","DOI":"10.2174\/138945007780618481","article-title":"The molecular basis of conantokin antagonism of NMDA receptor function","volume":"8","author":"Prorok","year":"2007","journal-title":"Curr. Drug Targets"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1111\/j.1476-5381.2011.01781.x","article-title":"Analgesic conotoxins: Block and G protein-coupled receptor modulation of N-type (Ca(V) 2.2) calcium channels","volume":"166","author":"Adams","year":"2012","journal-title":"Br. J. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"22747","DOI":"10.1074\/jbc.M115.648824","article-title":"Neurotoxins from snake venoms and \u03b1-conotoxin ImI inhibit functionally active ionotropic \u03b3-aminobutyric acid (GABA) receptors","volume":"290","author":"Kudryavtsev","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"582","DOI":"10.2174\/1381612822666151124234715","article-title":"Conotoxins: Structure, therapeutic potential and pharmacological applications","volume":"22","author":"Mir","year":"2016","journal-title":"Curr. Pharm. Des."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.peptides.2015.02.002","article-title":"Diversity of A-conotoxins of three worm-hunting cone snails (Conus brunneus, Conus nux, and Conus princeps) from the Mexican Pacific coast","volume":"68","author":"Aguilar","year":"2015","journal-title":"Peptides"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3259","DOI":"10.3390\/md13063259","article-title":"Structural and functional characterization of a novel \u03b1-Conotoxin Mr1.7 from Conus marmoreus targeting neuronal nAChR \u03b13\u03b22, \u03b19\u03b110 and \u03b16\/\u03b13\u03b22\u03b23 subtypes","volume":"13","author":"Wang","year":"2015","journal-title":"Mar. Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.1038\/s41598-017-18479-4","article-title":"A novel \u03b1-conopeptide Eu1.6 inhibits N-type (CaV2.2) calcium channels and exhibits potent analgesic activity","volume":"8","author":"Liu","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2004","DOI":"10.2174\/1381612822666160127112021","article-title":"Role of nicotinic and muscarinic receptors on synaptic plasticity and neurological diseases","volume":"22","author":"Fuenzalida","year":"2016","journal-title":"Curr. Pharm. Des."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.2174\/138161282210160304112133","article-title":"Targeting the nicotinic acetylcholine receptors (nAChRs) in astrocytes as a potential therapeutic target in Parkinson\u2019s disease","volume":"22","author":"Capani","year":"2016","journal-title":"Curr. Pharm. Des."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.pain.2010.09.007","article-title":"A novel mechanism of inhibition of high-voltage activated calcium channels by \u03b1-conotoxins contributes to relief of nerve injury-induced neuropathic pain","volume":"152","author":"Klimis","year":"2011","journal-title":"Pain"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"10233","DOI":"10.1074\/jbc.M110.196170","article-title":"Structure and activity of alpha-conotoxin PeIA at nicotinic acetylcholine receptor subtypes and GABA(B) receptor-coupled N-type calcium channels","volume":"286","author":"Daly","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4692","DOI":"10.1002\/anie.201600297","article-title":"Structure-activity studies of cysteine-rich \u03b1-conotoxins that inhibit high-voltage-activated calcium channels via GABA(B) receptor activation reveal a minimal functional motif","volume":"55","author":"Carstens","year":"2016","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.brainres.2005.08.009","article-title":"Alpha-conotoxin Vc1.1 alleviates neuropathic pain and accelerates functional recovery of injured neurons","volume":"1059","author":"Satkunanathan","year":"2005","journal-title":"Brain Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1016\/j.toxicon.2007.02.011","article-title":"From the identification of gene organization of alpha conotoxins to the cloning of novel toxins","volume":"49","author":"Yuan","year":"2007","journal-title":"Toxicon"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Giribaldi, J., and Dutertre, S. (2017). \u03b1-Conotoxins to explore the molecular, physiological and pathophysiological functions of neuronal nicotinic acetylcholine receptors. Neurosci. Lett.","DOI":"10.1016\/j.neulet.2017.11.063"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4226","DOI":"10.2174\/138161211798999384","article-title":"Structure-activity studies on alpha-conotoxins","volume":"17","author":"Muttenthaler","year":"2011","journal-title":"Curr. Pharm. Des."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1016\/j.bcp.2014.07.025","article-title":"Hydrophobic residues at position 10 of \u03b1-conotoxin PnIA influence subtype selectivity between \u03b17 and \u03b13\u03b22 neuronal nicotinic acetylcholine receptors","volume":"91","author":"Hopping","year":"2014","journal-title":"Biochem. Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.1021\/jm031010o","article-title":"Chemical and functional identification and characterization of novel sulfated alpha-conotoxins from the cone snail Conus anemone","volume":"47","author":"Loughnan","year":"2004","journal-title":"J. Med. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6628","DOI":"10.1021\/bi7004202","article-title":"Discovery, synthesis, and structure activity of a highly selective alpha7 nicotinic acetylcholine receptor antagonist","volume":"46","author":"Whiteaker","year":"2007","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"33610","DOI":"10.1074\/jbc.M205102200","article-title":"Alpha-conotoxin GIC from Conus geographus, a novel peptide antagonist of nicotinic acetylcholine receptors","volume":"277","author":"McIntosh","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3137","DOI":"10.1074\/jbc.M210280200","article-title":"Isolation, structure, and activity of GID, a novel alpha 4\/7-conotoxin with an extended N-terminal sequence","volume":"278","author":"Nicke","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1016\/j.bcp.2013.07.016","article-title":"Isolation and characterization of alpha-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors","volume":"86","author":"Inserra","year":"2013","journal-title":"Biochem. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1842","DOI":"10.1096\/fj.13-244103","article-title":"A novel \u03b14\/7-conotoxin LvIA from Conus lividus that selectively blocks \u03b13\u03b22 vs. \u03b16\/\u03b13\u03b22\u03b23 nicotinic acetylcholine receptors","volume":"28","author":"Luo","year":"2014","journal-title":"FASEB J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"7522","DOI":"10.1074\/jbc.271.13.7522","article-title":"A new alpha-conotoxin which targets alpha3beta2 nicotinic acetylcholine receptors","volume":"271","author":"Cartier","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"24678","DOI":"10.1074\/jbc.M602969200","article-title":"Alpha-conotoxin OmIA is a potent ligand for the acetylcholine-binding protein as well as alpha3beta2 and alpha7 nicotinic acetylcholine receptors","volume":"281","author":"Talley","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"30107","DOI":"10.1074\/jbc.M504102200","article-title":"A novel alpha-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between rat alpha9alpha10 and alpha7 nicotinic cholinergic receptors","volume":"280","author":"McIntosh","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2202","DOI":"10.1016\/j.neuropharm.2012.01.016","article-title":"Intrathecal \u03b1-conotoxins Vc1.1, AuIB and MII acting on distinct nicotinic receptor subtypes reverse signs of neuropathic pain","volume":"62","author":"Napier","year":"2012","journal-title":"Neuropharmacology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10943","DOI":"10.1523\/JNEUROSCI.3594-08.2008","article-title":"Analgesic \u03b1-conotoxins Vc1.1 and Rg1A inhibit N-Type calcium channels in rat sensory neurons via GABAB receptor activation","volume":"28","author":"Callaghan","year":"2008","journal-title":"J. Neurosci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.peptides.2017.06.008","article-title":"Cloning, expression and functional characterization of a D-superfamily conotoxin Lt28.1 with previously undescribed cysteine pattern","volume":"94","author":"Lu","year":"2017","journal-title":"Peptides"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.neuropharm.2015.08.047","article-title":"Molecular basis of toxicity of N-type calcium channel inhibitor MVIIA","volume":"101","author":"Wang","year":"2016","journal-title":"Neuropharmacology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.peptides.2016.04.004","article-title":"Im10A, a short conopeptide isolated from Conus imperialis and possesses two highly concentrated disulfide bridges and analgesic activity","volume":"81","author":"Yu","year":"2016","journal-title":"Peptides"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1124\/mol.114.096156","article-title":"Novel mechanism of voltage-gated N-type (Cav2.2) calcium channel inhibition revealed through \u03b1-conotoxin Vc1.1 activation of the GABA(B) receptor","volume":"87","author":"Huynh","year":"2015","journal-title":"Mol. Pharmacol."}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/16\/4\/112\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T14:59:11Z","timestamp":1760194751000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/16\/4\/112"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,3,31]]},"references-count":39,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2018,4]]}},"alternative-id":["md16040112"],"URL":"https:\/\/doi.org\/10.3390\/md16040112","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,3,31]]}}}