Understanding the role of protein arginylation in health and disease through analytical and biological approaches

• Grant number: 21/00140-3
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2021
• End date: April 03, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Giuseppe Palmisano
• Grantee: Janaína Macedo da Silva
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 18/15549-1 - Post-translational modifications in Chagas Disease biological processes and diagnostics: novel methodological approaches and biological applications, AP.JP2
• Associated scholarship(s): 22/15734-9 - Protein arginylation in amyotrophic lateral sclerosis (ALS): a link between ATE1 and TDP-43, BE.EP.DR

Abstract

Post-translational modifications (PTMs) alter the physicochemical and biological properties of a protein, influencing its function in health and disease state. Indeed, PTMs regulate stability, degradation, enzymatic function, protein interaction, and subcellular localization. Protein arginylation is a PTM little-explored described for the first time in 1963. Only in 2002, a study showed that the knockout of the ATE1 gene resulted in abnormalities in important processes such as cardiac development, angiogenesis, and tissue morphogenesis in mammals. Recently, our group identified arginylated proteins in Trypanosoma cruzi, however, the function of these PTMs in the pathogenesis is understood. In order to better understand this PTM, it is imperative to develop a method capable of analyzing and quantifying arginylated proteins, since at the moment there is no methodology in the literature for this purpose. Thus, the objective of this project is to develop and validate a robust, efficient, accurate, and reproducible enrichment method for the identification and quantification of arginylated peptides in complex biological matrices. For this, we will test different liquid chromatography formats from HPLC to microscale in-tip chromatographic columns. The enriched fractions will be analyzed by mass spectrometry. This method will be initially tested on synthetic arginylated peptides spiked in complex biological matrices. Once optimized, the method will be applied to T. cruzi epimastigote and trypomastigote life stages. Moreover, we will apply this method to LLCM-K2 rhesus monkey kidney cells infected and uninfected with T. cruzi. It should be noted that the method developed in this project will have a broad utility in the scientific community and will open the possibility to study this PTM in several biological mechanisms. Shedding lights on the identity of arginylated proteins is the most important step to understand the function and modulation of this PTM in pathophysiological conditions. Moreover, the levels and role of arginylation in biofluids is presently unknown and could introduce a new layer of protein regulation for biomarker discovery. (AU)

Scientific publications

(8)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARRASCO, ALEXIS GERMAN MURILLO; OTAKE, ANDREIA HANADA; MACEDO-DA-SILVA, JANAINA; SANTIAGO, VERONICA FEIJOLI; PALMISANO, GIUSEPPE; ANDRADE, LUCIANA NOGUEIRA DE SOUSA; CHAMMAS, ROGER. Deciphering the Functional Status of Breast Cancers through the Analysis of Their Extracellular Vesicles. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 24, n. 16, p. 33-pg., 2023-08-01. (18/15549-1, 18/18257-1, 20/04923-0, 21/00140-3, 19/05583-0)

MACEDO-DA-SILVA, JANAINA; ROSA-FERNANDES, LIVIA; GOMES, VINICIUS DE MORAIS; SANTIAGO, VERONICA FEIJOLI; SANTOS, DEIVID MARTINS; MOLNAR, CATARINA MARIA STANISCHESK; BARBOZA, BRUNO RAFAEL; DE SOUZA, EDMARCIA ELISA; MARQUES, RODOLFO FERREIRA; BOSCARDIN, SILVIA BEATRIZ; et al. Protein Arginylation Is Regulated during SARS-CoV-2 Infection. Viruses-Basel, v. 15, n. 2, p. 25-pg., 2023-02-01. (18/15549-1, 18/18257-1, 20/06409-1, 22/09915-0, 20/02988-7, 21/00140-3, 20/12277-0, 21/14179-9, 15/26722-8)

BARBOZA, BRUNO RAFAEL; MACEDO-DA-SILVA, JANAINA; SILVA, LAYS ADRIANNE MENDONCA TRAJANO; DE MORAIS GOMES, VINICIUS; SANTOS, DEIVID MARTINS; MARQUES-NETO, ANTONIO MOREIRA; MULE, SIMON NGAO; ANGELI, CLAUDIA BLANES; BORSOI, JULIANA; MORAES, CAROLINA BORSOI; et al. ST8Sia2 polysialyltransferase protects against infection by Trypanosoma cruzi. PLoS Neglected Tropical Diseases, v. 18, n. 9, p. 33-pg., 2024-09-01. (22/00796-9, 21/14751-4, 20/04923-0, 18/18257-1, 21/00140-3, 18/15549-1, 20/02988-7, 22/09915-0, 21/00507-4, 23/02096-7, 21/14179-9)

SAN FRANCISCO, JUAN; ASTUDILLO, CONSTANZA; LUIS VEGA, JOSE; CATALAN, ALEJANDRO; GUTIERREZ, BESSY; ARAYA, JORGE E.; ZAILBERGER, ANIBAL; MARINA, ANABEL; GARCIA, CARLOS; SANCHEZ, NURIA; et al. Trypanosoma cruzi pathogenicity involves virulence factor expression and upregulation of bioenergetic and biosynthetic pathways. VIRULENCE, v. 13, n. 1, p. 22-pg., 2022-12-31. (18/15549-1, 20/04923-0, 18/18257-1, 21/00140-3)

SANTIAGO, VERONICA FEIJOLI; DOMBROWSKI, JAMILLE GREGORIO; KAWAHARA, REBECA; ROSA-FERNANDES, LIVIA; MULE, SIMON NGAO; MURILLO, OSCAR; SANTANA, THAIS VIGGIANI; PACCINI COUTINHO, JOAO VICTOR; MACEDO-DA-SILVA, JANAINA; LAZARI, LUCAS CARDOSO; et al. Complement System Activation Is a Plasma Biomarker Signature during Malaria in Pregnancy. GENES, v. 14, n. 8, p. 20-pg., 2023-08-01. (21/14751-4, 18/15549-1, 20/06747-4, 20/04923-0, 17/04032-5, 21/00140-3, 19/12068-5, 18/20468-0)

MORETTI, ISABELE FATTORI; LERARIO, ANTONIO MARCONDES; SOLA, PAULA RODRIGUES; MACEDO-DA-SILVA, JANAINA; BAPTISTA, MAURICIO DA SILVA; PALMISANO, GIUSEPPE; OBA-SHINJO, SUELI MIEKO; MARIE, SUELY KAZUE NAGAHASHI. GBM Cells Exhibit Susceptibility to Metformin Treatment According to TLR4 Pathway Activation and Metabolic and Antioxidant Status. CANCERS, v. 15, n. 3, p. 18-pg., 2023-02-01. (20/04923-0, 18/18257-1, 04/12133-6, 20/02988-7, 21/00140-3, 13/07937-8)

CARNEIRO, ANDREIA; MACEDO-DA-SILVA, JANAINA; SANTIAGO, VERONICA FEIJOLI; DE OLIVEIRA, GILBERTO SANTOS; GUIMARAES, THIAGO; MENDONCA, CLARISSA FEROLLA; DE OLIVEIRA BRANQUINHO, JESSICA LAIS; LUCENA, CINTIA VERDAN; OSORIO, JULIANA; PERNAMBUCO, EDUARDO; et al. Urine proteomics as a non-invasive approach to monitor exertional rhabdomyolysis during military training. JOURNAL OF PROTEOMICS, v. 258, p. 17-pg., 2022-04-30. (18/13283-4, 14/27198-8, 18/18257-1, 20/04923-0, 18/15549-1, 21/00140-3)

LAZARI, LUCAS C.; DE OLIVEIRA, GILBERTO SANTOS; MACEDO-DA-SILVA, JANAINA; ROSA-FERNANDES, LIVIA; PALMISANO, GIUSEPPE. Mass spectrometry and machine learning in the identification of COVID-19 biomarkers. FRONTIERS IN ANALYTICAL SCIENCE, v. 3, p. 15-pg., 2023-03-31. (20/04923-0, 18/18257-1, 21/00140-3, 18/15549-1, 18/13283-4)