EVALUATION OF THE PRO-APOPTOTIC MECHANISM OF ACTION OF THE AMBLYOMIN-X

• Grant number: 10/07958-7
• Support type: Scholarships in Brazil - Doctorate
• Effective date (Start): July 01, 2010
• Effective date (End): May 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal researcher: Ana Marisa Chudzinski-Tavassi
• Grantee: Kátia Luciano Pereira Morais
• Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

The Amblyomin-X is a Kunitz-type inhibitor that was identified in the transcriptome of Amblyomma cajennense through analysis of ESTs sequences from a cDNA library, was expressed with a 6xHis-Tag fused at its N-terminus in E. coli, representing a protein with 13.4 kDa, with a single single chain protein that contains seven cysteine residues. This protein is is able to inhibit FXa activity in the presence of phospholipids and to prolong coagulation times overall as the activated partial thromboplastin time (APTT), prothrombin time (PT) and the PCA test - analysis of activity pro -coagulant. It was recently shown that the Amblyomin-X has pro-apoptotic activity in tumor cells and in vivo experiments, promotes regression of tumor mass and reduction of metastasis of some tumors tested. In vitro experiments showed that Amblyomin-X is able to inhibit the proteasome in tumor cells, increasing the pool of poly-ubiquitinated proteins, suggesting a possible disturbance of homeostasis of the endoplasmic reticulum (ER), which could be correlated with their pro-apoptotic effect. Thus, the purpose of this study is to investigate the internalization of Amblyomin-X by normal and tumor cells, as well as its structural state after this event and to correlate these results with the inhibition of proteasome caused by this protein, and to evaluate whether this effect promotes inhibition of ER stress, evaluating ways and marker proteins of this type of mechanism.

Scientific publications

(5)

(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)

CHUDZINSKI-TAVASSI, ANA MARISA; MORAIS, KATIA L. P.; FERNANDES PACHECO, MARIO THIEGO; MESQUITA PASQUALOTO, KERLY FERNANDA; DE SOUZA, JEAN GABRIEL. Tick salivary gland as potential natural source for the discovery of promising antitumor drug candidates. BIOMEDICINE & PHARMACOTHERAPY, v. 77, p. 14-19, FEB 2016. (10/07958-7, 98/14307-9, 13/07467-1, 11/05969-4)

MORAIS, KATIA L. P.; MESQUITA PASQUALOTO, KERLY FERNANDA; FERNANDES PACHECO, MARIO THIEGO; BERRA, CAROLINA MARIA; ALVAREZ-FLORES, MIRYAM PAOLA; CHUDZINSKI-TAVASSI, ANA MARISA. Rational development of a novel TFPI-like inhibitor from Amblyomma cajennense tick. Toxin Reviews, v. 33, n. 1-2, SI, p. 48-52, MAR-JUN 2014. (11/05969-4, 10/07958-7)

DE SOUZA, JEAN GABRIEL; MORAIS, KATIA L. P.; ANGLES-CANO, EDUARDO; BOUFLEUR, PAMELA; DE MELLO, EVANDRO SOBROZA; MARIA, DURVANEI AUGUSTO; TAEMI ORIGASSA, CLARICE SILVIA; ZAMPOLLI, HAMILTON DE CAMPOS; SARAIVA CAMARA, NIELS OLSEN; BERRA, CAROLINA MARIA; et al. Promising pharmacological profile of a Kunitz-type inhibitor in murine renal cell carcinoma model. ONCOTARGET, v. 7, n. 38, p. 62255-62266, SEP 20 2016. (12/06944-8, 10/07958-7, 12/02270-2, 13/07467-1, 15/50040-4)

PACHECO, MARIO T. F.; MORAIS, KATIA L. P.; BERRA, CAROLINA M.; DEMASI, MARILENE; SCIANI, JULIANA M.; BRANCO, VANIA G.; BOSCH, ROSEMARY V.; IQBAL, ASIF; CHUDZINSKI-TAVASSI, ANA MARISA. Specific role of cytoplasmic dynein in the mechanism of action of an antitumor molecule, Amblyomin-X. Experimental Cell Research, v. 340, n. 2, p. 248-258, JAN 15 2016. (11/05969-4, 13/07467-1, 10/07958-7)

MORAIS, KATIA L. P.; FERNANDES PACHECO, MARIO THIEGO; BERRA, CAROLINA MARIA; BOSCH, ROSEMARY V.; SCIANI, JULIANA MOZER; CHAMMAS, ROGER; SAITO, RENATA DE FREITAS; IQBAL, ASIF; CHUDZINSKI-TAVASSI, ANA MARISA. Amblyomin-X induces ER stress, mitochondrial dysfunction, and caspase activation in human melanoma and pancreatic tumor cell. Molecular and Cellular Biochemistry, v. 415, n. 1-2, p. 119-131, APR 2016. (10/07958-7, 98/14307-9, 13/07467-1, 11/05969-4)