{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T11:35:10Z","timestamp":1787657710436,"version":"build-2736575974"},"reference-count":50,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2023,7,26]],"date-time":"2023-07-26T00:00:00Z","timestamp":1690329600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hungarian Ministry for Innovation and Technology","award":["2018-1.2.1-NKP-2018-00005"],"award-info":[{"award-number":["2018-1.2.1-NKP-2018-00005"]}]},{"name":"Hungarian Ministry for Innovation and Technology","award":["RRF-2.3.1-21-2022-00015"],"award-info":[{"award-number":["RRF-2.3.1-21-2022-00015"]}]},{"name":"National Research, Development, and Innovation Fund of Hungary","award":["2018-1.2.1-NKP-2018-00005"],"award-info":[{"award-number":["2018-1.2.1-NKP-2018-00005"]}]},{"name":"National Research, Development, and Innovation Fund of Hungary","award":["RRF-2.3.1-21-2022-00015"],"award-info":[{"award-number":["RRF-2.3.1-21-2022-00015"]}]},{"name":"European Union\u2019s Recovery and Resilience Instrument","award":["2018-1.2.1-NKP-2018-00005"],"award-info":[{"award-number":["2018-1.2.1-NKP-2018-00005"]}]},{"name":"European Union\u2019s Recovery and Resilience Instrument","award":["RRF-2.3.1-21-2022-00015"],"award-info":[{"award-number":["RRF-2.3.1-21-2022-00015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Myostatin, an important negative regulator of muscle mass, is a therapeutic target for muscle atrophic disorders such as muscular dystrophy. Thus, the inhibition of myostatin presents a strategy to treat these disorders. It has long been established that the myostatin prodomain is a strong inhibitor of the mature myostatin, and the minimum peptide of the prodomain\u2014corresponding to the \u03b11-helix of its lasso-region\u2014responsible for the inhibitory efficiency was defined and characterized as well. Here we show that the minimum peptide segment based on the growth differentiation factor 11 (GDF11), which we found to be more helical in its stand-alone solvated stfate than the similar segment of myostatin, is a promising new base scaffold for inhibitor design. The proposed inhibitory peptides in their solvated state and in complex with the mature myostatin were analyzed by in silico molecule modeling supplemented with the electronic circular dichroism spectroscopy measurements. We defined the Gaussian\u2013Mahalanobis mean score to measure the fraction of dihedral angle-pairs close to the desired helical region of the Ramachandran-plot, carried out RING analysis of the peptide-protein interaction networks and characterized the internal motions of the complexes using our rigid-body segmentation protocol. We identified a variant\u201411m2\u2014that is sufficiently ordered both in solvent and within the inhibitory complex, forms a high number of contacts with the binding-pocket and induces such changes in its internal dynamics that lead to a rigidified, permanently locked conformation that traps this peptide in the binding site. We also showed that the naturally evolved \u03b11-helix has been optimized to simultaneously fulfill two very different roles: to function as a strong binder as well as a good leaving group. It forms an outstanding number of non-covalent interactions with the mature core of myostatin and maintains the most ordered conformation within the complex, while it induces independent movement of the gate-keeper \u03b2-hairpin segment assisting the dissociation and also results in the least-ordered solvated form which provides extra stability for the dissociated state and discourages rebinding.<\/jats:p>","DOI":"10.3390\/molecules28155655","type":"journal-article","created":{"date-parts":[[2023,7,27]],"date-time":"2023-07-27T02:07:17Z","timestamp":1690423637000},"page":"5655","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Inhibitor Design Strategy for Myostatin: Dynamics and Interaction Networks Define the Affinity and Release Mechanisms of the Inhibited Complexes"],"prefix":"10.3390","volume":"28","author":[{"given":"D\u00f3ra","family":"Nagy-Fazekas","sequence":"first","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"Hevesy Gy\u00f6rgy PhD School of Chemistry, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsolt","family":"Fazekas","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"Hevesy Gy\u00f6rgy PhD School of Chemistry, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"N\u00f3ra","family":"Taricska","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"ELKH-ELTE Protein Modeling Research Group, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P\u00e1l","family":"Str\u00e1ner","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"ELKH-ELTE Protein Modeling Research Group, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0095-5531","authenticated-orcid":false,"given":"D\u00f3ra","family":"Karancsin\u00e9 Menyh\u00e1rd","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"ELKH-ELTE Protein Modeling Research Group, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e1s","family":"Perczel","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Chemistry and Biology, Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"},{"name":"ELKH-ELTE Protein Modeling Research Group, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), Institute of Chemistry, E\u00f6tv\u00f6s Lor\u00e1nd University, P\u00e1zm\u00e1ny P\u00e9ter s\u00e9t\u00e1ny 1\/A, H-1117 Budapest, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/387083a0","article-title":"Regulation of Skeletal Muscle in Mice by a New TGF-Fl Member","volume":"3887","author":"McPherron","year":"1997","journal-title":"Nature"},{"key":"ref_2","unstructured":"LeRoith, D., and Bondy, C. (1996). A Multi-Volume Treatise, JAI Press."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"E866","DOI":"10.1073\/pnas.1714622115","article-title":"Molecular Characterization of Latent GDF8 Reveals Mechanisms of Activation","volume":"115","author":"Walker","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1544","DOI":"10.1021\/jm501170d","article-title":"Identification of the Minimum Peptide from Mouse Myostatin Prodomain for Human Myostatin Inhibition","volume":"58","author":"Takayama","year":"2015","journal-title":"J. Med. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1486","DOI":"10.1126\/science.1069525","article-title":"Induction of Cachexia in Mice by Systemically Administered Myostatin","volume":"296","author":"Zimmers","year":"2002","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1038\/nm.3512","article-title":"Transforming Growth Factor-\u03b2 Superfamily Ligand Trap ACE-536 Corrects Anemia by Promoting Late-Stage Erythropoiesis","volume":"20","author":"Suragani","year":"2014","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1038\/nature10152","article-title":"Latent TGF-\u03b2 Structure and Activation","volume":"474","author":"Shi","year":"2011","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"878069","DOI":"10.3389\/fendo.2022.878069","article-title":"Regulatory Role and Potential Importance of GDF-8 in Ovarian Reproductive Activity","volume":"13","author":"Zheng","year":"2022","journal-title":"Front. Endocrinol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1530\/REP-21-0197","article-title":"GDF-8 Stimulates Trophoblast Cell Invasion by Inducing ALK5-SMAD2\/3-Mediated MMP2 Expression","volume":"162","author":"Fang","year":"2021","journal-title":"Reproduction"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"9306","DOI":"10.1073\/pnas.151270098","article-title":"Regulation of Myostatin Activity and Muscle Growth","volume":"98","author":"Lee","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1007\/s13539-011-0035-5","article-title":"The Role of Myostatin in Muscle Wasting: An Overview","volume":"2","author":"Elkina","year":"2011","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"15842","DOI":"10.1073\/pnas.2534946100","article-title":"Activation of Latent Myostatin by the BMP-1tolloid Family of Metalloproteinases","volume":"100","author":"Wolfman","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"367","DOI":"10.15252\/embj.201797883","article-title":"Structure of the Human Myostatin Precursor and Determinants of Growth Factor Latency","volume":"37","author":"Cotton","year":"2018","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly Accurate Protein Structure Prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1074\/jbc.M111.270801","article-title":"Structure of Myostatin\u00b7follistatin-like 3: N-Terminal Domains of Follistatin-Type Molecules Exhibit Alternate Modes of Binding","volume":"287","author":"Cash","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1416","DOI":"10.1042\/BST0391416","article-title":"Biological Functions of the WAP Domain-Containing Multidomain Proteins WFIKKN1 and WFIKKN2","volume":"39","author":"Nagy","year":"2011","journal-title":"Biochem. Soc. Trans."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"23677","DOI":"10.1074\/jbc.M803025200","article-title":"Both WFIKKN1 and WFIKKN2 Have High Affinity for Growth and Differentiation Factors 8 and 11","volume":"283","author":"Trexler","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6333","DOI":"10.1074\/jbc.RA118.005831","article-title":"Crystal Structure of the WFIKKN2 Follistatin Domain Reveals Insight into How It Inhibits Growth Differentiation Factor 8 (GDF8) and GDF11","volume":"294","author":"McCoy","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1107\/S2053230X16001588","article-title":"Crystal Structure of Human GDF11","volume":"72","author":"Padyana","year":"2016","journal-title":"Acta Crystallogr. Sect. Struct. Biol. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Walker, R.G., Czepnik, M., Goebel, E.J., McCoy, J.C., Vujic, A., Cho, M., Oh, J., Aykul, S., Walton, K.L., and Schang, G. (2017). Structural Basis for Potency Differences between GDF8 and GDF11. BMC Biol., 15.","DOI":"10.1186\/s12915-017-0350-1"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.jchemneu.2018.02.003","article-title":"GDF11 Expression in the Adult Rat Central Nervous System","volume":"89","author":"Hayashi","year":"2018","journal-title":"J. Chem. Neuroanat."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.bbrc.2004.01.085","article-title":"Characterization and Identification of the Inhibitory Domain of GDF-8 Propeptide","volume":"315","author":"Jiang","year":"2004","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1002\/cmdc.201500533","article-title":"Effect of N-Terminal Acylation on the Activity of Myostatin Inhibitory Peptides","volume":"11","author":"Takayama","year":"2016","journal-title":"ChemMedChem"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Ohsawa, Y., Takayama, K., Nishimatsu, S.I., Okada, T., Fujino, M., Fukai, Y., Murakami, T., Hagiwara, H., Itoh, F., and Tsuchida, K. (2015). The Inhibitory Core of the Myostatin Prodomain: Its Interaction with Both Type I and II Membrane Receptors, and Potential to Treat Muscle Atrophy. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0133713"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1021\/acsmedchemlett.6b00420","article-title":"Structural Basis for the Effective Myostatin Inhibition of the Mouse Myostatin Prodomain-Derived Minimum Peptide","volume":"8","author":"Asari","year":"2017","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1021\/acsmedchemlett.7b00168","article-title":"Development of Potent Myostatin Inhibitory Peptides through Hydrophobic Residue-Directed Structural Modification","volume":"8","author":"Takayama","year":"2017","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1016\/j.bmc.2019.02.019","article-title":"Design and Synthesis of Potent Myostatin Inhibitory Cyclic Peptides","volume":"27","author":"Rentier","year":"2019","journal-title":"Bioorg. Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/j.cell.2010.07.011","article-title":"Reversal of Cancer Cachexia and Muscle Wasting by ActRIIB Antagonism Leads to Prolonged Survival","volume":"142","author":"Zhou","year":"2010","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/s13395-015-0060-8","article-title":"Myostatin Blockade with a Fully Human Monoclonal Antibody Induces Muscle Hypertrophy and Reverses Muscle Atrophy in Young and Aged Mice","volume":"5","author":"Latres","year":"2015","journal-title":"Skelet. Muscle"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1016\/S2213-8587(15)00298-3","article-title":"Myostatin Antibody (LY2495655) in Older Weak Fallers: A Proof-of-Concept, Randomised, Phase 2 Trial","volume":"3","author":"Becker","year":"2015","journal-title":"Lancet Diabetes Endocrinol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1021\/acs.oprd.0c00178","article-title":"Cost-Effective Flow Peptide Synthesis: Metamorphosis of HPLC","volume":"25","author":"Farkas","year":"2021","journal-title":"Org. Process Res. Dev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"147","DOI":"10.2174\/1389203716666151102105215","article-title":"Bacterial Expression and\/or Solid Phase Peptide Synthesis of 20-40 Amino Acid Long Polypeptides and Miniproteins, the Case Study of Class B GPCR Ligands","volume":"17","author":"Taricska","year":"2016","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/protein\/4.6.669","article-title":"Convex Constraint Analysis: A Natural Deconvolution of Circular Dichroism Curves of Proteins","volume":"4","author":"Perczel","year":"1991","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Percze1, A., Hollbsi, M., Foxman, B.M., and Fasman, G.D. (1991). Conformational Analysis of Pseudocyclic Hexapeptides Based on Quantitative Circular Dichroism (CD), NOE, and X-Ray Data. The Pure CD Spectra of Type I and Type I1@-Turns. J. Am. Chem. Soc., 113, 9772\u20139784.","DOI":"10.1021\/ja00026a010"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2937","DOI":"10.1111\/j.1432-1033.2004.04220.x","article-title":"The Optimization of Protein Secondary Structure Determination with Infrared and Circular Dichroism Spectra","volume":"271","author":"Oberg","year":"2004","journal-title":"Eur. J. Biochem."},{"key":"ref_36","unstructured":"(2023, June 25). ChirasCan Series AppliCation Note. Available online: https:\/\/technoinfo.ru\/technoinfo\/files\/012\/000\/0000012\/AppNote%20ACD%20Biosimilarity%20Studies%20of%20Serum%20Albumins.pdf."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1002\/psc.1169","article-title":"The Inherent Flexibility of Peptides and Protein Fragments Quantitized by CD in Conjunction with CCA+","volume":"15","author":"Perczel","year":"2009","journal-title":"J. Pept. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"W651","DOI":"10.1093\/nar\/gkac365","article-title":"RING 3.0: Fast Generation of Probabilistic Residue Interaction Networks from Structural Ensembles","volume":"50","author":"Clementel","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1968","DOI":"10.1002\/chem.201903826","article-title":"The Route from the Folded to the Amyloid State: Exploring the Potential Energy Surface of a Drug-Like Miniprotein","volume":"26","author":"Taricska","year":"2020","journal-title":"Chemistry"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1766","DOI":"10.1016\/j.str.2016.08.012","article-title":"A Structurally Dynamic Region of the HslU Intermediate Domain Controls Protein Degradation and ATP Hydrolysis","volume":"24","author":"Baytshtok","year":"2016","journal-title":"Structure"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.1080\/19420862.2016.1215786","article-title":"Beyond CDR-Grafting: Structure-Guided Humanization of Framework and CDR Regions of an Anti-Myostatin Antibody","volume":"8","author":"Apgar","year":"2016","journal-title":"MAbs"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2662","DOI":"10.1038\/emboj.2009.205","article-title":"The Structure of Myostatin:Follistatin 288: Insights into Receptor Utilization and Heparin Binding","volume":"28","author":"Cash","year":"2009","journal-title":"EMBO J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"5404","DOI":"10.1074\/jbc.RA119.012293","article-title":"Structural Basis of Specific Inhibition of Extracellular Activation of Pro- or Latent Myostatin by the Monoclonal Antibody SRK-015","volume":"295","author":"Dagbay","year":"2020","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3844","DOI":"10.1021\/acs.jcim.2c00397","article-title":"Omicron Binding Mode: Contact Analysis and Dynamics of the Omicron Receptor-Binding Domain in Complex with ACE2","volume":"62","author":"Fazekas","year":"2022","journal-title":"J. Chem. Inf. Model."},{"key":"ref_45","unstructured":"Abraham, L., and van der Spoel, H. (2021). GROMACS Documentation Release 2021.4, GROMACS Development Team."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"6269","DOI":"10.1021\/j100308a038","article-title":"The Missing Term in Effective Pair Potentialst","volume":"91","author":"Berendsen","year":"1987","journal-title":"J. Phys. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1002\/prot.24350","article-title":"Motional Timescale Predictions by Molecular Dynamics Simulations: Case Study Using Proline and Hydroxyproline Sidechain Dynamics","volume":"82","author":"Aliev","year":"2014","journal-title":"Proteins Struct. Funct. Bioinform."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1109\/MCSE.2007.55","article-title":"Matplotlib: A 2D Graphics Environment","volume":"9","author":"Hunter","year":"2007","journal-title":"Comput. Sci. Eng."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1589","DOI":"10.1093\/bioinformatics\/btg224","article-title":"PISCES: A Protein Sequence Culling Server","volume":"19","author":"Wang","year":"2003","journal-title":"Bioinformatics"},{"key":"ref_50","unstructured":"(2023, July 20). PyGraphViz. Available online: https:\/\/github.com\/pygraphviz\/pygraphviz."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/15\/5655\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:19:17Z","timestamp":1760127557000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/15\/5655"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,26]]},"references-count":50,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2023,8]]}},"alternative-id":["molecules28155655"],"URL":"https:\/\/doi.org\/10.3390\/molecules28155655","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,26]]}}}