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Are the interactions between collagen and proteins/enzymes present in the matriz vesicles responsible for the control in the biomineralization process?

Grant number: 14/00371-1
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): May 01, 2014
Effective date (End): October 31, 2017
Field of knowledge:Biological Sciences - Biochemistry - Enzymology
Principal Investigator:Pietro Ciancaglini
Grantee:Maytê Bolean Correia
Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated scholarship(s):15/06814-5 - Do interactions between collagen and proteins/enzymes present in matrix vesicles control biomineralization?, BE.EP.PD

Abstract

The biomineralization process occurs through the accumulation of mineral constituted mainly by phosphate and calcium ions which form the calcium phosphate salt with the structure of the hydroxiapatite. This process is mediated by osteoblasts, cells responsible for the beginning of the biomineralization process, mediated by release of the matrix vesicles (MVs). These vesicles arise from the cell's surface by budding and are secreted in the specific place in the beginning of biomineralization in the bone tissue matrix. MVs contain high concentrations of Ca2+ and inorganic phosphate (Pi) ions providing an adequate microenvironment for the initial formation and propagation of the hydroxiapatite crystals. Special attention should be given to some proteins present in the MVs: Annexin V (AnxA5), alkaline phosphatase (TNAP) and ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1). These proteins regulate the formation of calcium phosphate crystals, thus acting directly on the bone mineralization process. Among the annexins, the Annexin V specifically (protein with ~35 kDa) is responsible for the formation of calcium channel through the association of this protein with both inner and outer layer of the MVs membrane. The annexins are also responsible for the disorganization of cellular membrane resulting in the apoptosis process. TNAP is a nonspecific phosphomonohydrolase able to hydrolyze phosphate monoesters, pyrophosphate, phosphate diesters, as well as catalyze the transphosphorylation reactions. It's inserted in the plasmatic membrane of the MVs through the glycosylphosphatidylinositol anchor (GPI) and it is called "alkaline" for its ability to perform the substrate hydrolysis reactions more efficiently at pH above neutral (pH 8-11). Furthermore, TNAP has a crucial role in limiting the extracellular inorganic pyrophosphate concentration (ePPi), a potent inhibitor of mineralization, in order to maintain the Pi/PPi ratio adequate for a normal bone mineralization. The primary function of the TNAP is to degrade ePPi, which is produced extracellularly by NPP1. We intend to study the interactions of proteins (AnxA5, TNAP e NPP1), lipids and collagen (type II and X) in the biomineralization process, using the vesicular systems of proteoliposomes in order to form mimetic systems of MVs and study how these protein/protein and protein/collagen interactions can regulate and modulate the mineralization process mediated by MVs.

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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)
FAVARIN, B. Z.; BOLEAN, M.; RAMOS, A. P.; MAGRINI, A.; ROSATO, N.; MILLAN, J. L.; BOTTINI, M.; COSTA-FILHO, A. J.; CIANCAGLINI, P.. Lipid composition modulates ATP hydrolysis and calcium phosphate mineral propagation by TNAP-harboring proteoliposomes. Archives of Biochemistry and Biophysics, v. 691, . (17/08892-9, 14/00371-1, 18/12092-0, 15/06814-5)
BOTTINI, MASSIMO; MEBAREK, SAIDA; ANDERSON, KAREN L.; STRZELECKA-KILISZEK, AGNIESZKA; BOZYCKI, LUKASZ; SPER SIMAO, ANA MARIA; BOLEAN, MAYTE; CIANCAGLINI, PIETRO; PIKULA, JOANNA BANDOROWICZ; PIKULA, SLAWOMIR; et al. Matrix vesicles from chondrocytes and osteoblasts: Their biogenesis, properties, functions and biomimetic models. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1862, n. 3, p. 532-546, . (14/11941-3, 16/21236-0, 14/00371-1)
VESCHI, EKEVELINY AMABILE; BOLEAN, MAYTE; STRZELECKA-KILISZEK, AGNIESZKA; BANDOROWICZ-PIKULA, JOANNA; PIKULA, SLAWOMIR; GRANJON, THIERRY; MEBAREK, SAIDA; MAGNE, DAVID; RAMOS, ANA PAULA; ROSATO, NICOLA; et al. Localization of Annexin A6 in Matrix Vesicles During Physiological Mineralization. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 21, n. 4, . (17/08892-9, 14/00371-1, 15/06814-5)
SPER SIMAO, ANA MARIA; BOLEAN, MAYTE; FAVARIN, BRUNO ZOCCARATTO; VESCHI, EKEVELINY AMABILE; TOVANI, CAMILA BUSSOLA; RAMOS, ANA PAULA; BOTTINI, MASSIMO; BUCHET, RENE; MILLAN, JOSE LUIS; CIANCAGLINI, PIETRO. Lipid microenvironment affects the ability of proteoliposomes harboring TNAP to induce mineralization without nucleators. JOURNAL OF BONE AND MINERAL METABOLISM, v. 37, n. 4, p. 607-613, . (14/11941-3, 16/21236-0, 14/00371-1, 17/08892-9)
FAVARIN, B. Z.; ANDRADE, M. A. R.; BOLEAN, M.; SIMAO, A. M. S.; RAMOS, A. P.; HOYLAERTS, M. F.; MILLAN, J. L.; CIANCAGLINI, P.. Effect of the presence of cholesterol in the interfacial microenvironment on the modulation of the alkaline phosphatase activity during in vitro mineralization. COLLOIDS AND SURFACES B-BIOINTERFACES, v. 155, p. 466-476, . (16/21236-0, 15/00345-3, 12/20946-3, 14/11941-3, 14/00371-1)
GARCIA, A. F.; SIMAO, A. M. S.; BOLEAN, M.; HOYLAERTS, M. F.; MILLAN, J. L.; CIANCAGLINI, P.; COSTA-FILHO, A. J.. Effects of GPI-anchored TNAP on the dynamic structure of model membranes. Physical Chemistry Chemical Physics, v. 17, n. 39, p. 26295-26301, . (14/11941-3, 14/00371-1, 13/26088-1, 10/17662-8)
BOLEAN, MAYTE; BONIN, IVANA A.; SIMAO, ANA M. S.; BOTTINI, MASSIMO; BAGATOLLI, LUIS A.; HOYLAERTS, MARC F.; MILLAN, JOSE L.; CIANCAGLINI, PIETRO. Topographic analysis by atomic force microscopy of proteoliposomes matrix vesicle mimetics harboring TNAP and AnxA5. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, v. 1859, n. 10, p. 1911-1920, . (14/11941-3, 14/00371-1)
BOLEAN, MAYTE; IZZI, BENEDETTA; VAN KERCKHOVEN, SOETKIN; BOTTINI, MASSIMO; RAMOS, ANA PAULA; MILLAN, JOSE LUIS; HOYLAERTS, MARC F.; CIANCAGLINI, PIETRO. Matrix vesicle biomimetics harboring Annexin A5 and alkaline phosphatase bind to the native collagen matrix produced by mineralizing vascular smooth muscle cells. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1864, n. 8, . (17/08892-9, 14/00371-1, 15/06814-5)

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