{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T16:15:35Z","timestamp":1783440935531,"version":"3.54.6"},"reference-count":59,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T00:00:00Z","timestamp":1756857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Gyeongsangbuk-do (2025; Medicare Blue Food Development Project)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The potential of N-terminal mitochondrial-targeting sequences (MTSs) as potent antimicrobial peptides (AMPs) has been previously reported. Building on this, 3923 mitochondrial proteins were identified from various marine organisms, among which 470 MTSs were predicted using MitoFates. Of these, 25 MTSs were synthesized and assessed for antimicrobial activity. All MTSs exhibited antifungal activity against Candida albicans, while 22 and 20 MTSs demonstrated activity against Escherichia coli and Staphylococcus aureus, respectively. Notably, the MTS of methylcrotonyl-CoA carboxylase subunit 1 (MCCC1-MTS) derived from swimming crab (Portunus trituberculatus) and the MTS of dihydrolipoamide branched-chain transacylase E2 (DBT-MTS) derived from herring (Oncorhynchus keta) showed strong antimicrobial activity against both Gram-positive and Gram-negative bacteria, as well as fungi. In addition, MCCC1-MTS markedly reduced the viability of multiple cancer cell lines with minimal cytotoxicity toward HaCaT cells and effectively suppressed the growth of A549-xenografted tumors in BALB\/c nude mice without inducing weight loss. These findings demonstrate that MTSs derived from marine organisms function as potent AMPs with selective cytotoxicity toward cancer cells, further supporting previous evidence that protozoan MTSs represent novel AMP candidates.<\/jats:p>","DOI":"10.3390\/ijms26178546","type":"journal-article","created":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T08:04:15Z","timestamp":1756886655000},"page":"8546","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Marine-Derived N-Terminal Mitochondrial-Targeting Sequences Exhibit Antimicrobial and Anticancer Activities"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6142-8559","authenticated-orcid":false,"given":"Sun-Mee","family":"Hong","sequence":"first","affiliation":[{"name":"Department of Technology Development, Marine Industry Research Institute for East Sea Rim, Uljin 36315, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyu-Shik","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, College of Medicine, Dongguk University, Gyeongju 38766, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4318-1588","authenticated-orcid":false,"given":"Kyuho","family":"Jeong","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Dongguk University, Gyeongju 38766, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6183-3100","authenticated-orcid":false,"given":"Jongwan","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Anatomy, College of Medicine, Dongguk University, Gyeongju 38766, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4518-6537","authenticated-orcid":false,"given":"Eun-Young","family":"Yun","sequence":"additional","affiliation":[{"name":"Department of Integrative Bio-Industrial Engineering, Sejong University, Seoul 05006, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae Won","family":"Goo","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Dongguk University, Gyeongju 38766, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1042\/CS20220713","article-title":"Cancer and sepsis","volume":"137","author":"Williams","year":"2023","journal-title":"Clin. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1128\/AAC.03937-14","article-title":"Engineered cationic antimicrobial peptides to overcome multidrug resistance by ESKAPE pathogens","volume":"59","author":"Deslouches","year":"2015","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1517\/14712598.2013.844227","article-title":"Antimicrobial peptides: New drugs for bad bugs?","volume":"14","author":"Steckbeck","year":"2014","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2511","DOI":"10.1128\/AAC.02218-12","article-title":"Rational design of engineered cationic antimicrobial peptides consisting exclusively of arginine and tryptophan, and their activity against multidrug-resistant pathogens","volume":"57","author":"Deslouches","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"46635","DOI":"10.18632\/oncotarget.16743","article-title":"Antimicrobial peptides with selective antitumor mechanisms: Prospect for anticancer applications","volume":"8","author":"Deslouches","year":"2017","journal-title":"Oncotarget"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Gaspar, D., Veiga, A.S., and Castanho, M.A. (2013). From antimicrobial to anticancer peptides. A review. Front. Microbiol., 4.","DOI":"10.3389\/fmicb.2013.00294"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1016\/j.ijantimicag.2004.09.005","article-title":"Antimicrobial peptides: Premises and promises","volume":"24","author":"Reddy","year":"2004","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"20160153","DOI":"10.1098\/rsfs.2016.0153","article-title":"What can machine learning do for antimicrobial peptides, and what can antimicrobial peptides do for machine learning?","volume":"7","author":"Lee","year":"2017","journal-title":"Interface Focus"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/j.tibtech.2011.05.001","article-title":"The expanding scope of antimicrobial peptide structures and their modes of action","volume":"29","author":"Nguyen","year":"2011","journal-title":"Trends Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1016\/S0966-842X(00)01823-0","article-title":"The role of cationic antimicrobial peptides in innate host defences","volume":"8","author":"Hancock","year":"2000","journal-title":"Trends Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1016\/j.coph.2006.04.006","article-title":"Antibacterial peptides for therapeutic use: Obstacles and realistic outlook","volume":"6","author":"Marr","year":"2006","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_12","first-page":"2033","article-title":"Design of Synthetic Antimicrobial Peptides against Resistant Acinetobacter baumannii Using Computational Approach","volume":"8","year":"2017","journal-title":"Int. J. Pharm. Sci. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Mohamed, M.F., Abdelkhalek, A., and Seleem, M.N. (2016). Evaluation of short synthetic antimicrobial peptides for treatment of drug-resistant and intracellular Staphylococcus aureus. Sci. Rep., 6.","DOI":"10.1038\/srep29707"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3492","DOI":"10.1074\/jbc.M117.805499","article-title":"Computational antimicrobial peptide design and evaluation against multidrug-resistant clinical isolates of bacteria","volume":"293","author":"Nagarajan","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e329","DOI":"10.1002\/agt2.329","article-title":"The effect of tailing lipidation on the bioactivity of antimicrobial peptides and their aggregation tendency: Special issue: Emerging investigators","volume":"4","author":"Lin","year":"2023","journal-title":"Aggregate"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Grimsey, E., Collis, D.W., Mikut, R., and Hilpert, K. (2020). The effect of lipidation and glycosylation on short cationic antimicrobial peptides. Biochim. Biophys. Acta (BBA)-Biomembr., 1862.","DOI":"10.1016\/j.bbamem.2020.183195"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Armas, F., Di Stasi, A., Mardirossian, M., Romani, A.A., Benincasa, M., and Scocchi, M. (2021). Effects of lipidation on a proline-rich antibacterial peptide. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22157959"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.mib.2014.09.008","article-title":"Endosymbiotic theory for organelle origins","volume":"22","author":"Zimorski","year":"2014","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1002\/bies.20761","article-title":"Reconstructing evolution: Gene transfer from plastids to the nucleus","volume":"30","author":"Bock","year":"2008","journal-title":"Bioessays"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1016\/j.tcb.2016.07.001","article-title":"The Role of Charge in Protein Targeting Evolution","volume":"26","author":"Garg","year":"2016","journal-title":"Trends Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kunze, M., and Berger, J. (2015). The similarity between N-terminal targeting signals for protein import into different organelles and its evolutionary relevance. Front. Physiol., 6.","DOI":"10.3389\/fphys.2015.00259"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.cell.2009.08.005","article-title":"Importing mitochondrial proteins: Machineries and mechanisms","volume":"138","author":"Chacinska","year":"2009","journal-title":"Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1038\/s41580-020-0210-7","article-title":"The cell biology of mitochondrial membrane dynamics","volume":"21","author":"Giacomello","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.biochi.2022.07.011","article-title":"Biomembrane lipids: When physics and chemistry join to shape biological activity","volume":"203","year":"2022","journal-title":"Biochimie"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2400102","DOI":"10.1002\/adtp.202400102","article-title":"Mitochondrial N-Terminal Signal Sequences as Novel Antimicrobial and Annticancer Peptides","volume":"7","author":"Jo","year":"2024","journal-title":"Adv. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1074\/mcp.M114.043083","article-title":"MitoFates: Improved prediction of mitochondrial targeting sequences and their cleavage sites","volume":"14","author":"Fukasawa","year":"2015","journal-title":"Mol. Cell. Proteom."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Mol, A.R., Castro, M.S., and Fontes, W. (2018). NetWheels: A web application to create high quality peptide helical wheel and net projections. bioRxiv.","DOI":"10.1101\/416347"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"W449","DOI":"10.1093\/nar\/gkw329","article-title":"PEP-FOLD3: Faster de novo structure prediction for linear peptides in solution and in complex","volume":"44","author":"Lamiable","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2835","DOI":"10.1128\/AAC.04932-14","article-title":"Potential of Novel Antimicrobial Peptide P3 from Bovine Erythrocytes and Its Analogs To Disrupt Bacterial Membranes In Vitro and Display Activity against Drug-Resistant Bacteria in a Mouse Model","volume":"59","author":"Zhang","year":"2015","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1007\/s00284-014-0529-4","article-title":"In Vitro Synergistic Activities of Antimicrobial Peptide Brevinin-2CE with Five Kinds of Antibiotics Against Multidrug-Resistant Clinical Isolates","volume":"68","author":"Zhang","year":"2014","journal-title":"Curr. Microbiol."},{"key":"ref_31","first-page":"628","article-title":"Antibacterial activity of the recombinant antimicrobial peptide Ib-AMP4 from Impatiens balsamina and its synergy with other antimicrobial agents against drug resistant bacteria","volume":"68","author":"Fan","year":"2013","journal-title":"Pharmazie"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1016\/j.tube.2014.08.001","article-title":"Cationic amphipathic D-enantiomeric antimicrobial peptides with in vitro and ex vivo activity against drug-resistant Mycobacterium tuberculosis","volume":"94","author":"Lan","year":"2014","journal-title":"Tuberculosis"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Greco, I., Molchanova, N., Holmedal, E., Jenssen, H., Hummel, B., Watts, J., H\u00e5kansson, J., Hansen, P., and Svenson, J. (2020). Correlation between hemolytic activity, cytotoxicity and systemic in vivo toxicity of synthetic antimicrobial peptides. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-69995-9"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e01358-23","DOI":"10.1128\/msystems.01358-23","article-title":"PyAMPA: A high-throughput prediction and optimization tool for antimicrobial peptides","volume":"9","author":"Valle","year":"2024","journal-title":"mSystems"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Nikookar Golestani, R., Ghods, E., Rostamian, M., Madanchi, H., and Talebi, A.F. (2023). Investigating the antimicrobial activity, cytotoxicity, and action mechanism of acylated and amidated derivatives of AurH1 antifungal peptide. BMC Microbiol., 23.","DOI":"10.1186\/s12866-023-03090-7"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1016\/S0092-8674(00)81895-4","article-title":"Human \u03b2-defensin-1 is a salt-sensitive antibiotic in lung that is inactivated in cystic fibrosis","volume":"88","author":"Goldman","year":"1997","journal-title":"Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1046\/j.1523-1747.2001.01340.x","article-title":"Cutaneous injury induces the release of cathelicidin anti-microbial peptides active against group A Streptococcus","volume":"117","author":"Dorschner","year":"2001","journal-title":"J. Investig. Dermatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2845","DOI":"10.1128\/AAC.49.7.2845-2850.2005","article-title":"Antimicrobial and chemoattractant activity, lipopolysaccharide neutralization, cytotoxicity, and inhibition by serum of analogs of human cathelicidin LL-37","volume":"49","author":"Ciornei","year":"2005","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6896","DOI":"10.1128\/AAC.03628-14","article-title":"Effective control of Salmonella infections by employing combinations of recombinant antimicrobial human \u03b2-defensins hBD-1 and hBD-2","volume":"58","author":"Maiti","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1984","DOI":"10.1111\/febs.16276","article-title":"The impact of phosphatidylserine exposure on cancer cell membranes on the activity of the anticancer peptide HB43","volume":"289","author":"Antonietti","year":"2022","journal-title":"FEBS J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1789","DOI":"10.1007\/s00424-024-02948-7","article-title":"Phosphatidylserine externalization as immune checkpoint in cancer","volume":"476","author":"Kur","year":"2024","journal-title":"Pfl\u00fcgers Arch.-Eur. J. Physiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e45061","DOI":"10.7554\/eLife.45061","article-title":"The antimicrobial peptide defensin cooperates with tumour necrosis factor to drive tumour cell death in Drosophila","volume":"8","author":"Parvy","year":"2019","journal-title":"eLife"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s12934-022-01848-8","article-title":"The dual interaction of antimicrobial peptides on bacteria and cancer cells; mechanism of action and therapeutic strategies of nanostructures","volume":"21","author":"Parchebafi","year":"2022","journal-title":"Microb. Cell Factories"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"156","DOI":"10.15212\/bioi-2020-0013","article-title":"A review for antimicrobial peptides with anticancer properties: Re-purposing of potential anticancer agents","volume":"1","author":"Zhong","year":"2021","journal-title":"BIO Integr."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.2147\/IJN.S445333","article-title":"Anticancer mechanisms and potential anticancer applications of antimicrobial peptides and their nano agents","volume":"19","author":"Dong","year":"2024","journal-title":"Int. J. Nanomed."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Tornesello, A.L., Borrelli, A., Buonaguro, L., Buonaguro, F.M., and Tornesello, M.L. (2020). Antimicrobial peptides as anticancer agents: Functional properties and biological activities. Molecules, 25.","DOI":"10.3390\/molecules25122850"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.bbamcr.2012.05.029","article-title":"A reevaluation of dual-targeting of proteins to mitochondria and chloroplasts","volume":"1833","author":"Carrie","year":"2013","journal-title":"Biochim. Biophys. Acta-Mol. Cell Res."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Boto, A., P\u00e9rez de la Lastra, J., and Gonz\u00e1lez, C. (2018). The road from host-defense peptides to a new generation of antimicrobial drugs. Molecules, 23.","DOI":"10.3390\/molecules23020311"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Mahlapuu, M., Hakansson, J., Ringstad, L., and Bjorn, C. (2016). Antimicrobial Peptides: An Emerging Category of Therapeutic Agents. Front. Cell Infect Microbiol., 6.","DOI":"10.3389\/fcimb.2016.00194"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"678","DOI":"10.3892\/ijo.2020.5099","article-title":"Anticancer peptide: Physicochemical property, functional aspect and trend in clinical application","volume":"57","author":"Chiangjong","year":"2020","journal-title":"Int. J. Oncol."},{"key":"ref_51","first-page":"1011","article-title":"Physiological aspects of membrane lipid fluidity in malignancy","volume":"7","author":"Deliconstantinos","year":"1987","journal-title":"Anticancer Res."},{"key":"ref_52","first-page":"6132","article-title":"Increased exposure of anionic phospholipids on the surface of tumor blood vessels","volume":"62","author":"Ran","year":"2002","journal-title":"Cancer Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1593\/neo.101366","article-title":"Increased exposure of phosphatidylethanolamine on the surface of tumor vascular endothelium","volume":"13","author":"Stafford","year":"2011","journal-title":"Neoplasia"},{"key":"ref_54","first-page":"2569","article-title":"The glycosylation landscape of pancreatic cancer","volume":"17","author":"Munkley","year":"2019","journal-title":"Oncol. Lett."},{"key":"ref_55","first-page":"3919","article-title":"The antimicrobial peptides and their potential clinical applications","volume":"11","author":"Lei","year":"2019","journal-title":"Am. J. Transl. Res."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Li, C., Liu, H., Yang, Y., Xu, X., Lv, T., Zhang, H., Liu, K., Zhang, S., and Chen, Y. (2018). N-myristoylation of antimicrobial peptide CM4 enhances its anticancer activity by interacting with cell membrane and targeting mitochondria in breast cancer cells. Front. Pharmacol., 9.","DOI":"10.3389\/fphar.2018.01297"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/bs.mie.2020.04.044","article-title":"Cell-penetrating and mitochondrion-targeting molecules","volume":"Volume 641","author":"Kubi","year":"2020","journal-title":"Methods in Enzymology"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1907","DOI":"10.1128\/IAI.73.4.1907-1916.2005","article-title":"Apoptosis, pyroptosis, and necrosis: Mechanistic description of dead and dying eukaryotic cells","volume":"73","author":"Fink","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.ceb.2004.09.011","article-title":"Death by design: Apoptosis, necrosis and autophagy","volume":"16","author":"Edinger","year":"2004","journal-title":"Curr. Opin. Cell Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8546\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:38:22Z","timestamp":1760035102000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8546"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,3]]},"references-count":59,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["ijms26178546"],"URL":"https:\/\/doi.org\/10.3390\/ijms26178546","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,3]]}}}