Large Scale Discovery and De Novo-Assisted Sequencing of Cationic Antimicrobial Peptides (CAMPs) by Microparticle Capture and Electron-Transfer Dissociation (ETD) Mass Spectrometry.
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.
Melanie L
ML
Juba
Paul S
PS
Russo
Megan
M
Devine
Stephanie
S
Barksdale
Carlos
C
Rodriguez
Department of Biology, University of Florida , Gainesville, Florida 32611, United States.
Kent A
KA
Vliet
Joel M
JM
Schnur
Monique L
ML
van Hoek
Barney M
BM
Bishop
D000489:Alligators and Crocodiles / Q000097:blood*; D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000097:blood* / Q000494:pharmacology; D001409:Bacillus cereus / Q000187:drug effects / Q000254:growth & development; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D002853:Chromatography, Liquid; D055808:Drug Discovery; D004583:Electrons; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D020100:Hydrogels; D008826:Microbial Sensitivity Tests; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D010316:Particle Size; D040901:Proteomics / Q000295:instrumentation / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000254:growth & development; D020539:Sequence Analysis, Protein / Q000379:methods; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development; D053719:Tandem Mass Spectrometry / Q000379:methods*
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D020100:Hydrogels
PEAKS db; alligator; antimicrobial; de novo sequencing; de novo-assisted sequencing; electron transfer dissociation; mass spectrometry; microparticles; peptides