Characterization of macromolecular interactions of proteins involved in the selenocysteine synthesis from Escherichia coli

• Grant number: 12/23730-1
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: April 01, 2013
• End date: August 31, 2013
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Otavio Henrique Thiemann
• Grantee: Vitor Hugo Balasco Serrão
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated research grant: 08/57910-0 - National Institute of Structural Biotechnology and Medicinal Chemistry in Infectious Diseases, AP.TEM

Abstract

The investigation of the genetic code translation to proteins calls the interest for its central role in the cellular metabolism. In particular, the investigation of new amino acid synthesis, such as selenocystreine and pyrolysin that result in the expansion of the genetic code from the traditional 20 amino acid residues to a total of 22 residues.The selenocysteine (Sec, U) represents the major biological form of selenium and its incorporation is a co-translational process in response to a UGA in-frame stop codon and requires a complex enzymatic machinery to be accomplished.The complete set f genes involved in the selenocysteine incorporation in prokaryotes and eukaryotes is known, however, the molecular interactions between the different proteins has not been characterized.This project has the purpose of characterizing at the molecular and structural level the interactions between the Selenocysteine Synthetase (SELA) and the selenocysteine-specific Translation Elongation Factor with the ribosome in Escherichia coli.To achieve these purpose, fluorescence anisotropy and microcalorimetry techniques will be used for the determination of the interaction constants of these protein complexes. To obtain structural models of the protein complex Negative Stain Electron Microscopy, High Resolution Cryo-Electron Microscopy and Hydrogen/Deuterium Exchange Mass Spectroscopy (HDEX) will be employed. The proposed investigation will help to elucidate the mechanism of selencysteine incorporation in bacteria as well as in other domains of life. (AU)

Scientific publications

(16)

(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)

SCORTECCI, JESSICA FERNANDES; BALASCO SERRAO, VITOR HUGO; CHELESKI, JULIANA; TORINI, JULIANA ROBERTA; ROMANELLO, LARISSA; DEMARCO, RICARDO; PEREIRA, HUMBERTO D'MUNIZ. Spectroscopic and calorimetric assays reveal dependence on dCTP and two metals (Zn2+ + Mg2+) for enzymatic activity of Schistosoma mansoni deoxycytidylate (dCMP) deaminase. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 11, A, p. 1326-1335, NOV 2017. (12/05532-8, 12/23730-1, 12/14223-9, 12/15777-8)

GAVA, DANIELLE; BALASCO SERRAO, VITOR HUGO; FERNANDES, LANA TEIXEIRA; CANTAO, MAURICIO EGIDIO; CIACCI-ZANELLA, JANICE REIS; MORES, NELSON; SCHAEFER, REJANE. Structure analysis of capsid protein of Porcine circovirus type 2 from pigs with systemic disease. Brazilian Journal of Microbiology, v. 49, n. 2, p. 351-357, APR-JUN 2018. (12/23730-1)

DORES-SILVA, PAULO ROBERTO; RODRIGUES KIRALY, VANESSA THOMAZ; DE OLIVEIRA MORITZ, MILENE NOBREGA; BALASCO SERRAO, VITOR HUGO; SIQUEIRA DOS PASSOS, PATRICIA MARIA; SPAGNOL, VALENTINE; TEIXEIRA, FELIPE ROBERTI; GAVA, LISANDRA MARQUES; CAUVI, DAVID MARIO; INACIO RAMOS, CARLOS HENRIQUE; et al. New insights on human Hsp70-escort protein 1: Chaperone activity, interaction with liposomes, cellular localizations and HSPA's self-assemblies remodeling. International Journal of Biological Macromolecules, v. 182, p. 772-784, JUL 1 2021. (14/16646-0, 14/07206-6, 17/26131-5, 16/25798-3, 09/54216-9, 12/50161-8, 17/07879-9, 11/23110-0, 17/07335-9, 12/23730-1)

BALASCO SERRAO, VITOR HUGO; SILVA, IVAN ROSA; ALVES DA SILVA, MARCO TULIO; SCORTECCI, JSSICA FERNANDES; FERNANDES, ADRIANO DE FREITAS; THIEMANN, OTAVIO HENRIQUE. The unique tRNA(Sec) and its role in selenocysteine biosynthesis. Amino Acids, v. 50, n. 9, p. 1145-1167, SEP 2018. (11/24017-4, 16/20977-7, 13/17791-0, 12/23730-1, 10/04429-3)

TORINI, JULIANA ROBERTA; ROMANELLO, LARISSA; HELENO BATISTA, FERNANDA APARECIDA; BALASCO SERRAO, VITOR HUGO; FAHEEM, MUHAMMAD; ZERAIK, ANA ELIZA; BIRD, LOUISE; NETTLESHIP, JOANNE; REDDIVARI, YAMINI; OWENS, RAY; et al. The molecular structure of Schistosoma mansoni PNP isoform 2 provides insights into the nucleoside selectivity of PNPs. PLoS One, v. 13, n. 9, SEP 7 2018. (12/14223-9, 12/10213-9, 12/23730-1)

TORINI, JULIANA ROBERTA; FERNANDES, ADRIANO DE FREITAS; BALASCO SERRAO, VITOR HUGO; ROMANELLO, LARISSA; BIRD, LOUISE E.; NETTLESHIP, JOANNE E.; OWENS, RAYMOND J.; BRANDAO-NETO, JOSE; ZERAIK, ANA ELIZA; DEMARCO, RICARDO; et al. Characterization of a Schistosoma mansoni NDPK expressed in sexual and digestive organs. Molecular and Biochemical Parasitology, v. 231, JUL 2019. (12/14223-9, 13/20715-4, 12/10213-9, 12/23730-1)

BALASCO SERRAO, VITOR HUGO; TORINI DE SOUZA, JULIANA ROBERTA; ROMANELLO, LARISSA; PEREIRA, HUMBERTO D'MUNIZ. Schistosoma mansoni Purine and Pyrimidine Biosynthesis: Structures and Kinetic Experiments in the Search for the Best Therapeutic Target. CURRENT PHARMACEUTICAL DESIGN, v. 23, n. 45, p. 6967-6983, 2017. (12/05532-8, 12/14223-9, 12/10213-9, 12/23730-1)

BALASCO SERRAO, VITOR HUGO; ROMANELLO, LARISSA; CASSAGO, ALEXANDRE; TORINI DE SOUZA, JULIANA ROBERTA; CHELESKI, JULIANA; DEMARCO, RICARDO; BRANDAO-NETO, JOSE; PEREIRA, HUMBERTO D'MUNIZ. Structure and kinetics assays of recombinant Schistosoma mansoni dihydrofolate reductase. Acta Tropica, v. 170, p. 190-196, JUN 2017. (12/14223-9, 12/23730-1)

ZERAIK, ANA ELIZA; BALASCO SERRAO, VITOR HUGO; ROMANELLO, LARISSA; TORINI, JULIANA ROBERTA; CASSAGO, ALEXANDRE; DEMARCO, RICARDO; PEREIRA, HUMBERTO D'MUNIZ. Schistosoma mansoni displays an adenine phosphoribosyltransferase preferentially expressed in mature female gonads and vitelaria. Molecular and Biochemical Parasitology, v. 214, p. 82-86, JUN 2017. (12/10213-9, 12/05532-8, 12/14223-9, 13/20715-4, 12/23730-1)

SCORTECCI, JESSICA F.; SERRAO, VITOR HUGO B.; FERNANDES, ADRIANO F.; BASSO, LUIS G. M.; GUTIERREZ, RAISSA F.; ARAUJO, ANA PAULA U.; NETO, MARIO O.; THIEMANN, OTAVIO H.. Initial steps in selenocysteine biosynthesis: The interaction between selenocysteine lyase and selenophosphate synthetase. International Journal of Biological Macromolecules, v. 156, p. 18-26, AUG 1 2020. (12/23730-1, 16/20977-7, 14/00206-0, 13/17791-0, 14/16005-4)

ROMANELLO, LARISSA; ZERAIK, ANA ELIZA; FERNANDES, ADRIANO DE FREITAS; TORINI, JULIANA ROBERTA; BIRD, LOUISE E.; NETTLESHIP, JOANNE E.; RADA, HEATHER; REDDIVARI, YAMINI; OWENS, RAY J.; SERRAO, VITOR HUGO BALASCO; et al. In vitro and in vivo characterization of the multiple isoforms of Schistosoma mansoni hypoxanthine-guanine phosphoribosyltransferases. Molecular and Biochemical Parasitology, v. 229, p. 24-34, APR 2019. (12/05532-8, 12/14223-9, 13/20715-4, 12/23730-1)

FERNANDES, ADRIANO DE FREITAS; SERRAO, VITOR HUGO BALASCO; SCORTECCI, JESSICA FERNANDES; THIEMANN, OTAVIO HENRIQUE. Seryl-tRNA synthetase specificity for tRNA(sec) in Bacterial Sec biosynthesis. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1868, n. 8, AUG 2020. (16/20977-7, 12/23730-1, 14/16005-4)

ROMANELLO, LARISSA; BALASCO SERRAO, VITOR HUGO; TORINI, JULIANA ROBERTA; BIRD, LOUISE E.; NETTLESHIP, JOANNE E.; RADA, HEATHER; REDDIVARI, YAMINI; OWENS, RAY J.; DEMARCO, RICARDO; BRANDAO-NETO, JOSE; et al. Structural and kinetic analysis of Schistosoma mansoni Adenylosuccinate Lyase (SmADSL). Molecular and Biochemical Parasitology, v. 214, p. 27-35, JUN 2017. (12/05532-8, 12/14223-9, 12/23730-1)

GAVA, DANIELLE; BALASCO SERRAO, VITOR HUGO; FERNANDES, LANA TEIXEIRA; CANTAO, MAURICIO EGIDIO; CIACCI-ZANELLA, JANICE REIS; MORES, NELSON; SCHAEFER, REJANE. Structure analysis of capsid protein of Porcine circovirus type 2 from pigs with systemic disease. Brazilian Journal of Microbiology, v. 49, n. 2, p. 7-pg., 2018-04-01. (12/23730-1)

BALASCO SERRAO, VITOR HUGO; DE FREITAS FERNANDES, ADRIANO; MANSOR BASSO, LUIS GUILHERME; SCORTECCI, JESSICA FERNANDES; CRUSCA JUNIOR, EDSON; LOPES CORNELIO, MARINONIO; MONSON DE SOUZA, BIBIANA; PALMA, MARIO SERGIO; DE OLIVEIRA NETO, MARIO; THIEMANN, OTAVIO HENRIQUE. The Specific Elongation Factor to Selenocysteine Incorporation in Escherichia coli: Unique tRNA(Sec) Recognition and its Interactions. Journal of Molecular Biology, v. 433, n. 23, NOV 19 2021. (13/17791-0, 14/00206-0, 12/15777-8, 12/23730-1, 14/16005-4, 98/14138-2)

SCORTECCI, JESSICA FERNANDES; BALASCO SERRAO, VITOR HUGO; CHELESKI, JULIANA; TORINI, JULIANA ROBERTA; ROMANELLO, LARISSA; DEMARCO, RICARDO; PEREIRA, HUMBERTO D'MUNIZ. Spectroscopic and calorimetric assays reveal dependence on dCTP and two metals (Zn2+ + Mg2+) for enzymatic activity of Schistosoma mansoni deoxycytidylate (dCMP) deaminase. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 11, p. 10-pg., 2017-11-01. (12/05532-8, 12/14223-9, 12/15777-8, 12/23730-1)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SERRÃO, Vitor Hugo Balasco. Characterization of the macromolecular interactions of proteins involved in the synthesis of selenocysteines in Escherichia coli. 2017. Doctoral Thesis - Universidade de São Paulo (USP). Instituto de Física de São Carlos (IFSC/BT) São Carlos.