Synthesis, kinetic studies and applications of substrates and inhibitors for proteolytic enzymes

• Grant number: 12/50191-4
• Support Opportunities: Research Projects - Thematic Grants
• Start date: December 01, 2012
• End date: November 30, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Maria Aparecida Juliano
• Grantee: Maria Aparecida Juliano
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant(s): 18/13588-0 - Multi - user equipment approved in grant 2012/50191-4: mass spectrometer impact II - Bruker, AP.EMU
• Associated scholarship(s): 15/01829-4 - Human Tissue Kallikrein (KLK1) and human plasma kallikrein on metabolism of semaphorins and neurotrophins, BP.PD; 14/18877-9 - Proteolytic enzymes in skin physiology and pathophysiology, BP.PD; 14/18451-1 - Leptospira spp. strains database implemention by MALDI-TOF-MS and activity analysis of their secreted proteases, BP.IC; + associated scholarships 13/21455-6 - Applied study will bioanalytical search profile blood plasma amino acid carriers Syndrome Cri-Du-Chat, BP.MS; 13/07763-0 - Studies of Radical Acetylation of Proteins Using Proteomics Tools, BP.DR; 13/12106-8 - Screening and characterization of serine carboxyl peptidases and glutamic peptidases in bacteria isolated from the composting operation at the São Paulo Zoo Park, BP.PD; 13/06654-2 - Synthesis and functionalization of mesoporous silicas aiming application bioanalytical, BP.IC; 11/20941-9 - DEVELOPMENT AND USE OF FLUORESCENCE TECHNIQUES FOR THE STUDY OF PROTEIN CONFORMATIONAL CHANGES, BP.PD - associated scholarships

Abstract

Considerable progress in the study of proteases was based on the development of synthetic peptides, which resulted in the chromogenic and fluorescent substrates. Quenched fluorescent peptides based on the transfer of energy by resonance of a fluorescent to a quancher group was the more recent and significant progress in the field that we are involve since 1990's. The synthesis of these FRET (Fluorescence Resonance Energy Transfer) peptides in solid phase linked to flexible resins and in systems of parallel synthesis allowed us to study the substrate specificity of several proteases by systematic variations in all positions of peptides with up to 10 amino acids in length. The project presented here is based on the development of modern techniques of organic synthesis and protein biochemistry for study of proteases involved in physiological and pathological processes. The proposals of this project are: Synthesis and assays with proteases of FRET peptide libraries in solid phase and in solution. Synthesis of protease inhibitors using non-natural amino acids. Assays as protease inhibitors of tellurium organic compounds and nitroolefins, particularly the cysteine proteases. Systematic studies will be done with human kallikreins, Plamodium and Leishmania proteases, processing peptidases and oligopeptidases. Virus, fungi and bactéria proteases were also included. (AU)

Scientific publications

(31)

(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)

KARCHER, DANIEL; GRENFELL, RAFAELLA C.; MORENO, ANDREA MICKE; MORENO, LUISA ZANOLLI; VASCONCELLOS, SILVIO ARRUDA; HEINEMANN, MARCOS B.; DE ALMEIDA JUNIOR, JOAO N.; JULIANO, LUIZ; JULIANO, MARIA A.. Identification of pathogenic and nonpathogenic Leptospira species of Brazilian isolates by Matrix Assisted Laser Desorption/Ionization and Time Flight mass spectrometry. Brazilian Journal of Microbiology, v. 49, n. 4, p. 900-908, OCT-DEC 2018. (13/17136-2, 12/50191-4)

NANI, JOAO V.; LEE, RICHARD S.; YONAMINE, CAMILA M.; SANT'ANNA, OSVALDO A.; JULIANO, MARIA A.; GADELHA, ARY; MARI, JAIR J.; HAYASHI, MIRIAN A. F.. Evaluation of NDEL1 oligopeptidase activity in blood and brain in an animal model of schizophrenia: effects of psychostimulants and antipsychotics. SCIENTIFIC REPORTS, v. 10, n. 1, OCT 28 2020. (19/09207-3, 17/02413-1, 12/50191-4, 13/13392-4)

DO NASCIMENTO, EDILZA SILVA; ANAYA, KATYA; DE OLIVEIRA, JULIA MARIANO CAJU; DE LACERDA, JOSE THALLES JOCELINO GOMES; MILLER, MICHAEL EDWARD; DIAS, MERIELLEN; MENDES, MARIA ANITA; PALLONE, JULIANA DE AZEVEDO LIMA; ARNS, CLARICE WEIS; JULIANO, MARIA APARECIDA; et al. Identification of bioactive peptides released from in vitro gastrointestinal digestion of yam proteins (Dioscorea cayennensis). Food Research International, v. 143, MAY 2021. (12/50191-4, 18/13588-0)

MARCONDES, M. F. M.; ALVES, F. M.; ASSIS, D. M.; HIRATA, I. Y.; JULIANO, L.; OLIVEIRA, V.; JULIANO, M. A.. Substrate specificity of mitochondrial intermediate peptidase analysed by a support-bound peptide library. FEBS OPEN BIO, v. 5, p. 429-436, 2015. (12/50191-4, 11/20941-9, 14/00661-0)

OLIVEIRA, JULIANA R.; BERTOLIN, THIAGO C.; ANDRADE, DOUGLAS; OLIVEIRA, LILIAN C. G.; KONDO, MARCIA Y.; SANTOS, JORGE A. N.; BLABER, MICHAEL; JULIANO, LUIZ; SEVERINO, BEATRICE; CALIENDO, GIUSEPPE; et al. Specificity studies on Kallikrein-related peptidase 7 (KLK7) and effects of osmolytes and glycosaminoglycans on its peptidase activity. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1854, n. 1, p. 73-83, JAN 2015. (12/50191-4)

STEINBERGER, JUTTA; GRISHKOVSKAYA, IRINA; CENCIC, REGINA; JULIANO, LUIZ; JULIANO, MARIA A.; SKERN, TIM. Foot-and-mouth disease virus leader proteinase: Structural insights into the mechanism of intermolecular cleavage. VIROLOGY, v. 468, p. 397-408, NOV 2014. (12/50191-4)

OKAMOTO, DEBORA N.; KONDO, MARCIA Y.; OLIVEIRA, LILIAN C. G.; HONORATO, RODRIGO V.; ZANPHORLIN, LETICIA M.; CORONADO, MONIKA A.; ARAUJO, MARIANA S.; DA MOTTA, GUACYARA; VERONEZ, CAMILA L.; ANDRADE, SHEILA S.; et al. P-I class metalloproteinase from Bothrops moojeni venom is a post-proline cleaving peptidase with kininogenase activity: Insights into substrate selectivity and kinetic behavior. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1844, n. 3, p. 545-552, MAR 2014. (12/50191-4)

ABREU, AFONSO G.; FRAGA, TATIANA R.; GRANADOS MARTINEZ, ADRIANA P.; KONDO, MARCIA Y.; JULIANO, MARIA A.; JULIANO, LUIZ; NAVARRO-GARCIA, FERNANDO; ISAAC, LOURDES; BARBOSA, ANGELA S.; ELIAS, WALDIR P.. The Serine Protease Pic From Enteroaggregative Escherichia coli Mediates Immune Evasion by the Direct Cleavage of Complement Proteins. Journal of Infectious Diseases, v. 212, n. 1, p. 106-115, JUL 1 2015. (12/50191-4, 11/14103-0, 10/50043-0)

ORLANDI-MATTOS, PAULO EDUARDO; AGUIAR, RODRIGO BARBOSA; VAZ JUNIOR, ITABAJARA DA SILVA; MORAES, JANE ZVEITER; DE ARAUJO CARLINI, ELISALDO LUIZ; JULIANO, MARIA APARECIDA; JULIANO, LUIZ. Enkephalin related peptides are released from jejunum wall by orally ingested bromelain. Peptides, v. 115, p. 32-42, MAY 2019. (12/50191-4)

REISS-PISTILLI, MARIA L. V.; SCHUPPAN, DETLEF; BARROSO, MADALENA M. S.; ASSUNCAO-MIRANDA, IRANAIA; FARIAS, SHIRLEY; LERY, LETICIA; BAUER, MICHAEL; JULIANO, LUIZ; JULIANO, MARIA A.; COELHO-SAMPAIO, TATIANA. An extracellular proteasome releases endostatin from human collagen XVIII. ANGIOGENESIS, v. 20, n. 1, p. 125-137, FEB 2017. (12/50191-4)

JOCELINO GOMES DE LACERDA, JOSE THALLES; RODRIGUES E LACERDA, RODRIGO; ASSUNCAO, NILSON ANTONIO; TASHIMA, ALEXANDRE KEIJI; JULIANO, MARIA APARECIDA; DOS SANTOS, JR., GILBERTO ALVES; DE SOUZA, MILENY DOS SANTOS; BATISTA, JACINTO DE LUNA; ROSSI, CARLOS EDUARDO; DE ALMEIDA GADELHA, CARLOS ALBERTO; et al. New insights into lectin from Abelmoschus esculentus seeds as a Kunitz-type inhibitor and its toxic effects on Ceratitis capitata and root-knot nematodes Meloidogyne spp.. Process Biochemistry, v. 63, p. 96-104, DEC 2017. (12/19321-9, 12/02514-9, 12/50191-4)

SONG, EUN SUK; JANG, HYEIN; GUO, HOU-FU; JULIANO, MARIA A.; JULIANO, LUIZ; MORRIS, ANDREW J.; GALPERIN, EMILIA; RODGERS, DAVID W.; HERSH, LOUIS B.. Inositol phosphates and phosphoinositides activate insulin-degrading enzyme, while phosphoinositides also mediate binding to endosomes. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 114, n. 14, p. E2826-E2835, APR 4 2017. (12/50191-4)

CARVALHAES, CECILIA GODOY; RAMOS, ANA CAROLINA; GONCALVES DE OLIVEIRA, LILIAN CAROLINE; JULIANO, MARIA APARECIDA; GALES, ANA CRISTINA. Rapid detection of ceftazidime/avibactam resistance by MALDI-TOF MS. Journal of Antimicrobial Chemotherapy, v. 73, n. 9, p. 2579-2582, SEP 2018. (12/50191-4)

DE SOUZA, EDUARDO SERGIO; KATAGIRI, ALBERTO H.; JULIANO, LUIZ; JULIANO, MARIA APARECIDA; PIMENTA, DANIEL CARVALHO; ITO, AMANDO SIUITI. Fret Studies of Conformational Changes in Heparin-Binding Peptides. Journal of Fluorescence, v. 24, n. 3, p. 885-894, MAY 2014. (12/50191-4)

OLIVEIRA, LILIAN C. G.; OKAMOTO, DEBORA N.; OLIVEIRA, JULIANA R.; KONDO, MARCIA Y.; GOUVEA, IURI E.; BITEAU, NICOLAS; BALTZ, THEO; MURAKAMI, MARIO T.; JULIANO, LUIZ; JULIANO, MARIA A.. Analysis of peptidase activities of a cathepsin B-like (TcoCBc1) from Trypanosoma congolense. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1844, n. 7, p. 1260-1267, JUL 2014. (12/50191-4)

RAMOS, PATRICIA L.; KONDO, MARCIA Y.; SANTOS, SAARA M. B.; DE VASCONCELLOS, SUZAN P.; ROCHA, RAFAEL C. S.; DA CRUZ, JOAO B.; EUGENIO, PATRICIA F. M.; CABRAL, HAMILTON; JULIANO, MARIA A.; JULIANO, LUIZ; et al. A Tropical Composting Operation Unit at SAo Paulo Zoo as a Source of Bacterial Proteolytic Enzymes. Applied Biochemistry and Biotechnology, v. 187, n. 1, p. 282-297, JAN 2019. (14/07037-0, 12/50191-4, 13/12106-8, 11/06548-2)

SEVERINO, BEATRICE; FIORINO, FERDINANDO; CORVINO, ANGELA; CALIENDO, GIUSEPPE; SANTAGADA, VINCENZO; ASSIS, DIEGO MAGNO; OLIVEIRA, JULIANA R.; JULIANO, LUIZ; MANGANELLI, SERENA; BENFENATI, EMILIO; et al. Synthesis, biological evaluation, and docking studies of PAR2-AP-derived pseudopeptides as inhibitors of kallikrein 5 and 6. Biological Chemistry, v. 396, n. 1, p. 45-52, JAN 2015. (12/50191-4)

SILVA, ROBERTA N.; OLIVEIRA, LILIAN C. G.; PARISE, CAROLINA B.; OLIVEIRA, JULIANA R.; SEVERINO, BEATRICE; CORVINO, ANGELA; DI VAIO, PAOLA; TEMUSSI, PIERO A.; CALIENDO, GIUSEPPE; SANTAGADA, VINCENZO; et al. Activity of human kallikrein-related peptidase 6 (KLK6) on substrates containing sequences of basic amino acids. Is it a processing protease?. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 5, p. 558-564, MAY 2017. (12/50191-4)

ASSIS, DIEGO M.; JULIANO, LUIZ; PASCHOALIN, THAYSA; KOUYOUMDJIAN, MARIA; CALIXTO, JOAO B.; SANTOS, ROBSON A. S.; PERTINHEZ, THELMA A.; GAUTHIER, FRANCIS; MOREAU, THIERRY; BLABER, MICHAEL; et al. Pharmacological Activities and Hydrolysis by Peptidases of [Phospho-Ser(6)]-Bradykinin (pS(6)-BK). Biochemical Pharmacology, v. 97, n. 2, p. 203-214, SEP 15 2015. (12/50191-4)

HAMIN NETO, YOUSSEF A. A.; DE OLIVEIRA, LILIAN C. G.; DE OLIVEIRA, ARTHUR H. C.; ROSA, JOSE C.; JULIANO, MARIA A.; JULIANO, LUIZ; RODRIGUES, ANDRE; CABRAL, HAMILTON. Determination of Specificity and Biochemical Characteristics of Neutral Protease Isolated from Myceliophthora thermophila. PROTEIN AND PEPTIDE LETTERS, v. 22, n. 11, p. 972-982, 2015. (12/50191-4, 12/18278-2, 11/06986-0, 12/24703-8)

PEREIRA, FELIPE V.; FERREIRA-GUIMARAES, CARLA A.; PASCHOALIN, THAYSA; SCUTTI, JORGE A. B.; MELO, FILIPE M.; SILVA, LUIS S.; MELO, AMANDA C. L.; SILVA, PRISCILA; TIAGO, MANOELA; MATSUO, ALISSON L.; et al. A Natural Bacterial-Derived Product, the Metalloprotease Arazyme, Inhibits Metastatic Murine Melanoma by Inducing MMP-8 Cross-Reactive Antibodies. PLoS One, v. 9, n. 4, APR 30 2014. (12/50191-4, 10/51423-0)

RIBEIRO GOMES MAIA, ANA CAROLINA; PORCINO, GABRIANE NASCIMENTO; FARIA-PINTO, PRISCILA; MENDES, TULIO VIEIRA; RIBEIRO ANTINARELLI, LUCIANA MARIA; COIMBRA, ELAINE SOARES; REIS, ALEXANDRE BARBOSA; JULIANO, LUIZ; JULIANO, MARIA APARECIDA; MARQUES, MARCOS JOSE; et al. Leishmania infantum nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1) B-domain: Antibody antiproliferative effect on the promastigotes and IgG subclass responses in canine visceral leishmaniasis. Veterinary Parasitology, v. 271, p. 38-44, JUL 2019. (12/50191-4)

OLIVEIRA, JOCELIA P. C.; FREITAS, RENATO F.; DE MELO, LEANDRO SILVA; BARROS, THALITA G.; SANTOS, JORGE A. N.; JULIANO, MARIA A.; PINHEIRO, SERGIO; BLABER, MICHAEL; JULIANO, LUIZ; MURI, ESTELA M. F.; et al. Isomannide-Based Peptidomimetics as Inhibitors for Human Tissue Kallikreins 5 and 7. ACS Medicinal Chemistry Letters, v. 5, n. 2, p. 128-132, FEB 2014. (12/50191-4, 11/51297-8)

KONDO, MARCIA Y.; GOUVEA, IURI E.; OKAMOTO, DEBORA N.; SANTOS, JORGE A. N.; SOUCCAR, CADEN; ODA, KOHEI; JULIANO, LUIZ; JULIANO, MARIA A.. Analysis of catalytic properties of tripeptidyl peptidase I (TTP-I), a serine carboxyl lysosomal protease, and its detection in tissue extracts using selective FRET peptide substrate. Peptides, v. 76, p. 80-86, FEB 2016. (13/12106-8, 12/50191-4)

DE OLIVEIRA, TALITA MENDES; JOCELINO GOMES DE LACERDA, JOSE THALLES; FIGUEIREDO LEITE, GIUSEPPE GIANINI; DIAS, MERIELLEN; MENDES, MARIA ANITA; KASSAB, PAULO; SALES E SILV, CLISLEYD GLAUCIELLY; JULIANO, MARIA APARECIDA; FORONES, NORA MANOUKIAN. Label-free peptide quantification coupled with in silico mapping of proteases for identification of potential serum biomarkers in gastric adenocarcinoma patients. CLINICAL BIOCHEMISTRY, v. 79, p. 61-69, MAY 2020. (18/13588-0, 12/50191-4)

BEZERRA, TALIANA; ESTEVEZ, MARIO; LACERDA, JOSE THALLES; DIAS, MERIELLEN; JULIANO, MARIA; MENDES, MARIA ANITA; MORGANO, MARCELO; PACHECO, MARIA TERESA; MADRUGA, MARTA. Chicken Combs and Wattles as Sources of Bioactive Peptides: Optimization of Hydrolysis, Identification by LC-ESI-MS2 and Bioactivity Assessment. Molecules, v. 25, n. 7, APR 2020. (18/13588-0, 12/50191-4)

INDARTE, MARTIN; LAZZA, CRISTIAN M.; ASSIS, DIEGO; CAFFINI, NESTOR O.; JULIANO, MARIA A.; AVILES, FRANCESC X.; DAURA, XAVIER; LOPEZ, LAURA M. I.; TREJO, SEBASTIAN A.. A Bowman-Birk protease inhibitor purified, cloned, sequenced and characterized from the seeds of Maclura pomifera (Raf.) Schneid. PLANTA, v. 245, n. 2, p. 343-353, FEB 2017. (12/50191-4)

ALENCAR BEZERRA, TALIANA KENIA; JOCELINO GOMES DE LACERDA, JOSE THALLES; SALU, BRUNO RAMOS; VILELA OLIVA, MARIA LUIZA; JULIANO, MARIA APARECIDA; PACHECO, MARIA TERESA BERTOLDO; MADRUGA, MARTA SUELY. Identification of Angiotensin I-Converting Enzyme-Inhibitory and Anticoagulant Peptides from Enzymatic Hydrolysates of Chicken Combs and Wattles. JOURNAL OF MEDICINAL FOOD, v. 22, n. 12, DEC 2019. (18/13588-0, 12/50191-4)

GILIO, JOYCE M.; MARCONDES, MARCELO F.; FERRARI, DEBORA; JULIANO, MARIA A.; JULIANO, LUIZ; OLIVEIRA, VITOR; MACHADO, MAURICIO F. M.. Processing of metacaspase 2 from Trypanosoma brucei (TbMCA2) broadens its substrate specificity. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 4, p. 388-394, APR 2017. (15/11190-0, 14/20847-0, 12/50191-4)

DAMEN, MARK; IZIDORO, MARIO A.; OKAMOTO, DEBORA N.; OLIVEIRA, LILIAN C. G.; AMATDJAIS-GROENEN, HELENE I., V; VAN CLEEF, KOEN W. R.; VAN RIJ, RONALD P.; DIETEREN, CINDY E. J.; GIRONES, DANIEL; VAN BUUREN, BERND N. M.; et al. Cationic Geminoid Peptide Amphiphiles Inhibit DENV2 Protease, Furin, and Viral Replication. Molecules, v. 27, n. 10, p. 15-pg., 2022-05-01. (12/50191-4)

TELES, CATHERINE M.; ANTUNES, VICTOR U.; CARDOSO, RAQUEL S.; CANDIDO, TUANY Z.; LIMA, CARMEN S. P.; RUIZ, ANA LUCIA T. G.; JULIANO, MARIA A.; FAVARO, DENIZE C.; ABBEHAUSEN, CAMILLA. Functionalization of new anticancer Pt(II) complex with transferrin receptor binding peptide. Inorganica Chimica Acta, v. 511, OCT 1 2020. (19/16904-2, 16/07729-4, 12/50191-4)

Filed patent(s) as a result of this research project