Large Scale Discovery and De Novo-Assisted Sequencing of Cationic Antimicrobial Peptides (CAMPs) by Microparticle Capture and Electron-Transfer Dissociation (ETD) Mass SpectrometryClick to copy article linkArticle link copied!
- Melanie L. Juba
- Paul S. Russo
- Megan Devine
- Stephanie Barksdale
- Carlos Rodriguez
- Kent A. Vliet
- Joel M. Schnur
- Monique L. van Hoek
- Barney M. Bishop
Abstract

The identification and sequencing of novel cationic antimicrobial peptides (CAMPs) have proven challenging due to the limitations associated with traditional proteomics methods and difficulties sequencing peptides present in complex biomolecular mixtures. We present here a process for large-scale identification and de novo-assisted sequencing of newly discovered CAMPs using microparticle capture followed by tandem mass spectrometry equipped with electron-transfer dissociation (ETD). This process was initially evaluated and verified using known CAMPs with varying physicochemical properties. The effective parameters were then applied in the analysis of a complex mixture of peptides harvested from American alligator plasma using custom-made (Bioprospector) functionalized hydrogel particles. Here, we report the successful sequencing process for CAMPs that has led to the identification of 340 unique peptides and the discovery of five novel CAMPs from American alligator plasma.
Cited By
Smart citations by scite.ai include citation statements extracted from the full text of the citing article. The number of the statements may be higher than the number of citations provided by ACS Publications if one paper cites another multiple times or lower if scite has not yet processed some of the citing articles.
This article is cited by 26 publications.
- Xiaolei Bian, Zhifu Chen, Fang Li, Yuanyuan Xie, Yi Li, Youhong Luo, Xiangman Zou, Hui Wang, Jingwen Zhang, Xiaowen Wang, Jinyong Zhang, Dongliang Guan. Single Amine or Guanidine Modification on Norvancomycin and Vancomycin to Overcome Multidrug-Resistance through Augmented Lipid II Binding and Increased Membrane Activity. Journal of Medicinal Chemistry 2024, 67
(22)
, 20639-20663. https://doi.org/10.1021/acs.jmedchem.4c02196
- Nicholas M. Riley and Joshua J. Coon . The Role of Electron Transfer Dissociation in Modern Proteomics. Analytical Chemistry 2018, 90
(1)
, 40-64. https://doi.org/10.1021/acs.analchem.7b04810
- Barney M. Bishop, Melanie L. Juba, Paul S. Russo, Megan Devine, Stephanie M. Barksdale, Shaylyn Scott, Robert Settlage, Pawel Michalak, Kajal Gupta, Kent Vliet, Joel M. Schnur, and Monique L. van Hoek . Discovery of Novel Antimicrobial Peptides from Varanus komodoensis (Komodo Dragon) by Large-Scale Analyses and De-Novo-Assisted Sequencing Using Electron-Transfer Dissociation Mass Spectrometry. Journal of Proteome Research 2017, 16
(4)
, 1470-1482. https://doi.org/10.1021/acs.jproteome.6b00857
- Praveen Nagella, Balamuralikrishnan Balasubramanian, Sungkwon Park, Udisha Singh, Arpita Jayan, Saptadeepa Mukherjee, Aatika Nizam, Arun Meyyazhagan, Manikantan Pappuswamy, Joseph Kadanthottu Sebastian, Vasantha Veerappa Lakshmaiah, Amin Mousavi Khaneghah. Production, Delivery, and Regulatory Aspects for Application of Plant-Based Anti-microbial Peptides: a Comprehensive Review. Probiotics and Antimicrobial Proteins 2025, 22 https://doi.org/10.1007/s12602-024-10421-1
- Angela Michela Immacolata Montone, Sara Elsa Aita, Federico Capuano, Angelo Citro, Alessandra Esposito, Alfonso Gallo, Morena Nappa, Enrico Taglioni, Carmela Maria Montone. Detection of antibacterial peptides in artisanal rennet and evaluation of their antibacterial activity. International Dairy Journal 2024, 57 , 106074. https://doi.org/10.1016/j.idairyj.2024.106074
- Ashley M. Carpenter, Monique L. van Hoek. Development of a defibrinated human blood hemolysis assay for rapid testing of hemolytic activity compared to computational prediction. Journal of Immunological Methods 2024, 529 , 113670. https://doi.org/10.1016/j.jim.2024.113670
- Monique L. van Hoek, Fahad M. Alsaab, Ashley M. Carpenter. GATR-3, a Peptide That Eradicates Preformed Biofilms of Multidrug-Resistant Acinetobacter baumannii. Antibiotics 2024, 13
(1)
, 39. https://doi.org/10.3390/antibiotics13010039
- Mengmeng Zhang, Shiwei Li, Jianjun Zhao, Quan Shuang, Yanan Xia, Fengmei Zhang. A novel endogenous antimicrobial peptide MP-4 derived from koumiss of Inner Mongolia by peptidomics, and effects on Staphylococcus aureus. LWT 2024, 191 , 115595. https://doi.org/10.1016/j.lwt.2023.115595
- Ali Andalibi, Remi Veneziano, Mikell Paige, Michael Buschmann, Amanda Haymond, Virginia Espina, Alessandra Luchini, Lance Liotta, Barney Bishop, Monique Van Hoek. Drug discovery efforts at George Mason University. SLAS Discovery 2023, 28
(6)
, 270-274. https://doi.org/10.1016/j.slasd.2023.03.001
- Ulka Gawde, Shuvechha Chakraborty, Faiza Hanif Waghu, Ram Shankar Barai, Ashlesha Khanderkar, Rishikesh Indraguru, Tanmay Shirsat, Susan Idicula-Thomas. CAMPR4: a database of natural and synthetic antimicrobial peptides. Nucleic Acids Research 2023, 51
(D1)
, D377-D383. https://doi.org/10.1093/nar/gkac933
- Liana Chafran, Amy Carfagno, Amaal Altalhi, Barney Bishop. Green Hydrogel Synthesis: Emphasis on Proteomics and Polymer Particle-Protein Interaction. Polymers 2022, 14
(21)
, 4755. https://doi.org/10.3390/polym14214755
- Amanda R. P. Silva, Marina S. Guimarães, Jheniffer Rabelo, Lisandra Herrera Belén, Caio José Perecin, Jorge G Farías, João H. P. M. Santos, Carlota O. Rangel-Yagui. Recent advances in the design of antimicrobial peptide conjugates. Journal of Materials Chemistry B 2022, 10
(19)
, 3587-3600. https://doi.org/10.1039/D1TB02757C
- Randall R. Jiménez, Amy Carfagno, Luke Linhoff, Brian Gratwicke, Douglas C. Woodhams, Liana Soares Chafran, Molly C. Bletz, Barney Bishop, Carly R. Muletz-Wolz, . Inhibitory Bacterial Diversity and Mucosome Function Differentiate Susceptibility of Appalachian Salamanders to Chytrid Fungal Infection. Applied and Environmental Microbiology 2022, 88
(8)
https://doi.org/10.1128/aem.01818-21
- Albert T. Lebedev, Irina D. Vasileva, Tatiana Y. Samgina. FT‐MS in the de novo top‐down sequencing of natural nontryptic peptides. Mass Spectrometry Reviews 2022, 41
(2)
, 284-313. https://doi.org/10.1002/mas.21678
- Megan E. Duffy, Jacquelyn A. Neibauer, Jamee Adams, Rachel A. Lundeen, Gabrielle Rocap, Anitra E. Ingalls, Clara A. Fuchsman, Richard G. Keil. Protein cycling in the eastern tropical North Pacific oxygen‐deficient zone: A de novo‐discovery peptidomic approach. Limnology and Oceanography 2022, 67
(2)
, 498-510. https://doi.org/10.1002/lno.12012
- Maria Magana, Muthuirulan Pushpanathan, Ana L Santos, Leon Leanse, Michael Fernandez, Anastasios Ioannidis, Marc A Giulianotti, Yiorgos Apidianakis, Steven Bradfute, Andrew L Ferguson, Artem Cherkasov, Mohamed N Seleem, Clemencia Pinilla, Cesar de la Fuente-Nunez, Themis Lazaridis, Tianhong Dai, Richard A Houghten, Robert E W Hancock, George P Tegos. The value of antimicrobial peptides in the age of resistance. The Lancet Infectious Diseases 2020, 20
(9)
, e216-e230. https://doi.org/10.1016/S1473-3099(20)30327-3
- Faiza Hanif Waghu, Susan Idicula‐Thomas. Collection of antimicrobial peptides database and its derivatives: Applications and beyond. Protein Science 2020, 29
(1)
, 36-42. https://doi.org/10.1002/pro.3714
- Monique L. van Hoek, M. Dennis Prickett, Robert E. Settlage, Lin Kang, Pawel Michalak, Kent A. Vliet, Barney M. Bishop. The Komodo dragon (Varanus komodoensis) genome and identification of innate immunity genes and clusters. BMC Genomics 2019, 20
(1)
https://doi.org/10.1186/s12864-019-6029-y
- Jessica R. Shartouny, Joshy Jacob. Mining the tree of life: Host defense peptides as antiviral therapeutics. Seminars in Cell & Developmental Biology 2019, 88 , 147-155. https://doi.org/10.1016/j.semcdb.2018.03.001
- Xiaoyan Hou, Shanshan Li, Qingying Luo, Guanghui Shen, Hejun Wu, Meiliang Li, Xingyan Liu, Anjun Chen, Meng Ye, Zhiqing Zhang. Discovery and identification of antimicrobial peptides in Sichuan pepper (Zanthoxylum bungeanum Maxim) seeds by peptidomics and bioinformatics. Applied Microbiology and Biotechnology 2019, 103
(5)
, 2217-2228. https://doi.org/10.1007/s00253-018-09593-y
- Sergey V. Kovalev, Albert T. Lebedev. Identification of biologically active peptides by means of Fourier transform mass spectrometry. 2019, 425-468. https://doi.org/10.1016/B978-0-12-814013-0.00014-4
- Megan Devine, Melanie Juba, Paul Russo, Barney Bishop. Structurally stable N-t-butylacrylamide hydrogel particles for the capture of peptides. Colloids and Surfaces B: Biointerfaces 2018, 161 , 471-479. https://doi.org/10.1016/j.colsurfb.2017.11.001
- Ezra M. C. Chung, Scott N. Dean, Crystal N. Propst, Barney M. Bishop, Monique L. van Hoek. Komodo dragon-inspired synthetic peptide DRGN-1 promotes wound-healing of a mixed-biofilm infected wound. npj Biofilms and Microbiomes 2017, 3
(1)
https://doi.org/10.1038/s41522-017-0017-2
- Stephanie M. Barksdale, Evelyn J. Hrifko, Monique L. van Hoek. Cathelicidin antimicrobial peptide from Alligator mississippiensis has antibacterial activity against multi-drug resistant Acinetobacter baumanii and Klebsiella pneumoniae. Developmental & Comparative Immunology 2017, 70 , 135-144. https://doi.org/10.1016/j.dci.2017.01.011
- Osmel Fleitas Martinez, Caleb Mawuli Agbale, Fernanda Nomiyama, Octávio Luiz Franco. Deciphering bioactive peptides and their action mechanisms through proteomics. Expert Review of Proteomics 2016, 13
(11)
, 1007-1016. https://doi.org/10.1080/14789450.2016.1238305
- Monique L. van Hoek. Diversity in Host Defense Antimicrobial Peptides. 2016, 3-26. https://doi.org/10.1007/978-3-319-32949-9_1
Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.
Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.
The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.