Proteomics
The structural analysis of peptides and proteins, and the identification of unknown proteins are among the most important aspects of biotechnology for both basic and applied research. The Proteomic and Mass Spectrometry Facility at CEINGE provides a series of high technological analytical services addressed to the structural characterization of peptides, proteins and glycoproteins based on the integration of classical biochemical procedures with advanced mass spectrometric methodologies. These analytical services are grouped in the following categories:
- Mass Spectrometry Services
- Proteomic Services
All analytical services include:
Appropriate scientific consultance for customers; Interpretation of the results; Storage of data for at least three years; Data conversion to publication quality forms. A scientific report on the analytical data signed by the Responsible of the Facility is available on request.
Special agreement can be stipulated for large numbers of samples.
When complex Services not precisely fitting the standard list are needed, the most suitable strategies to approach the problem will be identified and individual quotations will be provided according to the specific needs.
Contact:
Prof. Maria Monti montimar@unina.it; phone 0039 0813737919
Dr. Simona Celentano celentano@ceinge.unina.it; phone 0039 0813737895
Mass Spectrometry Services
Accurate Molecular Weight determination
Determination of the accurate molecular mass of peptides and proteins by either Electrospray or MALDI mass spectrometric analyses. Desalting of the samples by HPLC is also included.
Peptide mapping, post-translational modifications and variants.
Complete characterisation of native and recombinant proteins including analysis of protein variants and mutants, identification of post-translational modifications and proteolytic processing, and assignemnt of S-S bridges by MALDI mapping. Pre-treatment of the sample either in solution or in gel and enzymatic hydrolysis are also included.
Peptide sequencing by tandem MS
Direct analysis of purified samples by tandem mass spectrometry experiments using either MALDI-TOF/TOF or ES-MS/MS techniques. When needed, HPLC separation of peptides is also performed.
Analysis of Glycoproteins
Complete characterisation of the oligosaccharide moieties in glycoproteins. The Service includes: Protein digestion, enzymatic stripping of glycoforms, MALDI/MS profiling of glycoforms, structural determination of individual glycoforms by MALDI-TOF/TOF techniques.
Sugar analysis
Determination of the monosaccharide composition of glycoproteins by chemical hydrolysis, sugar derivatization and GC-MS analysis.
Analysis of metabolytes
Pre-treatment of the sample, chemical derivatization and GC-MS analysis or direct investigation by LC-MS/MS techniques in MRM scan mode. Quantitative determination of metabolytes can also be performed on request.
Proteomic Services
Identification of proteins by LCMS/MS
Identification of proteins either in solution opr following fractionation by 1D or 2D electrophoretic methodologies. Excission of protein band(s), in situ digestion, LC-MS/MS analysis of the peptide mixtures, Database Search by Mascot software.
Preparation of protein samples for identification by SDS-PAGE
Separation of protein samples by SDS-PAGE for identification of protein band(s)
Quantitative Differential Proteomic Analyses
Identification and quantification of proteins differentially expressed in cell lines or tissues by tandem mass spectrometry methodologies according to the label free approach. The entire protein contents is extracted from samples, purified by solid phase extraction, digested with trypsin and subjected to LC-MS/MS analysis. Label free quantification of the differentially expressed proteins is performed by evaluating the current intensity associated with each peptide according to the extracted ionic method (XIC) or the number of fragmentation observed for each peptide ion (spectral counts, SpC).
PAPERS
Spettrometria di massa
S-Glutathionylation at Cys328 and Cys542 Impairs STAT3 Phosphorylation | ACS Chemical Biology
June 18, 2014 -ACS Chemical Biology
S-Glutathionylation at Cys328 and Cys542 Impairs STAT3 Phosphorylation
Elena Butturini, Elena Darra, Giulia Chiavegato, Barbara Cellini, Flora Cozzolino, Maria Monti, Piero Pucci, Daniele Dell’Orco, and Sofia Mariotto
S-glutathionylation exerts opposing roles in the regulation of STAT1 and STAT3 signaling in reactive microglia - ScienceDirect
February 3, 2018 -Free Radical Biology and Medicine
S-glutathionylation exerts opposing roles in the regulation of STAT1 and STAT3 signaling in reactive microglia
Elena Butturini, Flora Cozzolino, Diana Boriero, Alessandra Carcereri de Prati, Maria Monti, Michele Rossin, Diana Canetti, Barbara Cellini, Piero Pucci, Sofia Mariotto
Proteomica
Lysosome purinergic receptor P2X4 regulates neoangiogenesis induced by microvesicles from sarcoma patients | Cell Death & Disease (nature.com)
August 17, 2021 -Cell Death & Disease
Lysosome purinergic receptor P2X4 regulates neoangiogenesis induced by microvesicles from sarcoma patients
Wulf Palinski, Maria Monti, Rosa Camerlingo, Ilaria Iacobucci, Serena Bocella, Federica Pinto, Clara Iannuzzi, Gelsomina Mansueto, Sara Pignatiello, Flavio Fazioli, Michele Gallo, Laura Marra, Flora Cozzolino, Annarosaria De Chiara, Piero Pucci, Antonio Bilancio & Filomena de Nigris
Biomolecules | Free Full-Text | What Antarctic Plants Can Tell Us about Climate Changes: Temperature as a Driver for Metabolic Reprogramming (mdpi.com)
July 23, 2021-Biomolecules
What Antarctic Plants Can Tell Us about Climate Changes: Temperature as a Driver for Metabolic Reprogramming
Laura Bertini, Flora Cozzolino, Silvia Proietti, Gaia Salvatore Falconieri, Ilaria Iacobucci, Rosanna Salvia, Patrizia Falabella, Maria Monti, Carla Caruso
Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis | Acta Neuropathologica Communications | Full Text (biomedcentral.com)
May 3, 2021-Acta Neuropathologica Communications
Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis
Kevin Peikert, Enrica Federti, Alessandro Matte, Gabriela Constantin, Enrica Caterina Pietronigro, Paolo Francesco Fabene, Paola Defilippi, Emilia Turco, Federico Del Gallo, Pietro Pucci, Angela Amoresano, Anna Illiano, Flora Cozzolino, Maria Monti, Francesca Garello, Enzo Terreno, Seth Leo Alper, Hannes Glaß, Lisann Pelzl, Katja Akgün, Tjalf Ziemssen, Rainer Ordemann, Florian Lang, Anna Maria Brunati, Lucia De Franceschi
Proteome alterations in erythrocytes with PIEZO1 gain-of-function mutations | Blood Advances | American Society of Hematology (ashpublications.org)
JANUARY 3, 2023-blood advances
Proteome alterations in erythrocytes with PIEZO1 gain-of-function mutations
Immacolata Andolfo, Vittoria Monaco, Flora Cozzolino, Barbara Eleni Rosato, Roberta Marra, Vincenza Cerbone, Valeria Maria Pinto, Gian Luca Forni, Sule Unal, Achille Iolascon, Maria Monti, Roberta Russo