Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis

L-carnosine (Car) is an endogenous dipeptide with significant potential in drug discovery for neurodegenerative diseases, while TAT1, a small arginine-rich peptide derived from the HIV-1 trans-activator protein (TAT), is known to stimulate proteasome activity. In this study, three isomeric peptides...

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Main Authors: Alicia Maroto, Olivier Briand, Alessia Distefano, Filiz Arioz, Olivier Monasson, Elisa Peroni, Giuseppe Grasso, Christine Enjalbal, Antony Memboeuf
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Language:English
Published: MDPI AG 2025-02-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/4/853
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author Alicia Maroto
Olivier Briand
Alessia Distefano
Filiz Arioz
Olivier Monasson
Elisa Peroni
Giuseppe Grasso
Christine Enjalbal
Antony Memboeuf
author_facet Alicia Maroto
Olivier Briand
Alessia Distefano
Filiz Arioz
Olivier Monasson
Elisa Peroni
Giuseppe Grasso
Christine Enjalbal
Antony Memboeuf
author_sort Alicia Maroto
collection DOAJ
description L-carnosine (Car) is an endogenous dipeptide with significant potential in drug discovery for neurodegenerative diseases, while TAT1, a small arginine-rich peptide derived from the HIV-1 trans-activator protein (TAT), is known to stimulate proteasome activity. In this study, three isomeric peptides were synthesised by incorporating the Car moiety at the N-terminus, C-terminus, or central position of the TAT1 sequence. To differentiate these isomers, high-resolution and energy-resolved CID MS/MS experiments were conducted. The resulting MS/MS spectra showed a high degree of similarity among the peptides, predominantly characterised by fragment ion peaks arising from arginine-specific neutral losses. Energetic analysis was similarly inconclusive in resolving the isomers. However, Principal Component Analysis (PCA) enabled clear differentiation of the three peptides by considering the entire MS/MS spectra rather than focusing solely on precursor ion intensities or major fragment peaks. PCA loadings revealed distinct fragment ions for each peptide, albeit with lower intensities, providing insights into consecutive fragmentation patterns. Some of these specific peaks could also be attributed to scrambling during fragmentation. These results demonstrate the potential of PCA as a simple chemometric tool for semi-automated peak identification in complex MS/MS spectra.
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spelling doaj-art-3571182d85644b73a6c54fc1829d3f8f2025-08-20T03:12:12ZengMDPI AGMolecules1420-30492025-02-0130485310.3390/molecules30040853Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component AnalysisAlicia Maroto0Olivier Briand1Alessia Distefano2Filiz Arioz3Olivier Monasson4Elisa Peroni5Giuseppe Grasso6Christine Enjalbal7Antony Memboeuf8Univ Brest, CEMCA, CNRS, UMR 6521, 29238 Brest, FranceUniv Brest, CEMCA, CNRS, UMR 6521, 29238 Brest, FranceUniv Brest, CEMCA, CNRS, UMR 6521, 29238 Brest, FranceUniv Brest, CEMCA, CNRS, UMR 6521, 29238 Brest, FranceCY Cergy Paris Université, CNRS, BioCIS, 95000 Cergy Pontoise, FranceCY Cergy Paris Université, CNRS, BioCIS, 95000 Cergy Pontoise, FranceChemical Sciences Department, University of Catania, 95125 Catania, ItalyUniv Montpellier, CNRS, ENSCM, IBMM, 34093 Montpellier, FranceUniv Brest, CEMCA, CNRS, UMR 6521, 29238 Brest, FranceL-carnosine (Car) is an endogenous dipeptide with significant potential in drug discovery for neurodegenerative diseases, while TAT1, a small arginine-rich peptide derived from the HIV-1 trans-activator protein (TAT), is known to stimulate proteasome activity. In this study, three isomeric peptides were synthesised by incorporating the Car moiety at the N-terminus, C-terminus, or central position of the TAT1 sequence. To differentiate these isomers, high-resolution and energy-resolved CID MS/MS experiments were conducted. The resulting MS/MS spectra showed a high degree of similarity among the peptides, predominantly characterised by fragment ion peaks arising from arginine-specific neutral losses. Energetic analysis was similarly inconclusive in resolving the isomers. However, Principal Component Analysis (PCA) enabled clear differentiation of the three peptides by considering the entire MS/MS spectra rather than focusing solely on precursor ion intensities or major fragment peaks. PCA loadings revealed distinct fragment ions for each peptide, albeit with lower intensities, providing insights into consecutive fragmentation patterns. Some of these specific peaks could also be attributed to scrambling during fragmentation. These results demonstrate the potential of PCA as a simple chemometric tool for semi-automated peak identification in complex MS/MS spectra.https://www.mdpi.com/1420-3049/30/4/853isomeric peptidescarnosineTAT1high-resolution mass spectrometryPrincipal Component AnalysisMS/MS
spellingShingle Alicia Maroto
Olivier Briand
Alessia Distefano
Filiz Arioz
Olivier Monasson
Elisa Peroni
Giuseppe Grasso
Christine Enjalbal
Antony Memboeuf
Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
Molecules
isomeric peptides
carnosine
TAT1
high-resolution mass spectrometry
Principal Component Analysis
MS/MS
title Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
title_full Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
title_fullStr Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
title_full_unstemmed Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
title_short Differentiation of Isomeric TAT1-CARNOSINE Peptides by Energy-Resolved Mass Spectrometry and Principal Component Analysis
title_sort differentiation of isomeric tat1 carnosine peptides by energy resolved mass spectrometry and principal component analysis
topic isomeric peptides
carnosine
TAT1
high-resolution mass spectrometry
Principal Component Analysis
MS/MS
url https://www.mdpi.com/1420-3049/30/4/853
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