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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Lipid composition modulates ATP hydrolysis and calcium phosphate mineral propagation by TNAP-harboring proteoliposomes

Texto completo
Autor(es):
Favarin, B. Z. [1, 2] ; Bolean, M. [2] ; Ramos, A. P. [2] ; Magrini, A. [3] ; Rosato, N. [4] ; Millan, J. L. [5] ; Bottini, M. [5, 4] ; Costa-Filho, A. J. [1] ; Ciancaglini, P. [2]
Número total de Autores: 9
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Dept Phys, FFCLRP, Ribeirao Preto, SP - Brazil
[2] Univ Sao Paulo, FFCLRP, Dept Chem, Ribeirao Preto, SP - Brazil
[3] Univ Roma Tor Vergata, Dept Biopathol & Imaging Diagnost, Rome - Italy
[4] Univ Roma Tor Vergata, Dept Expt Med, Rome - Italy
[5] Sanford Burnham Prebys Med Discovery Inst, La Jolla, CA - USA
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: Archives of Biochemistry and Biophysics; v. 691, SEP 30 2020.
Citações Web of Science: 0
Resumo

Bone biomineralization is mediated by a special class of extracellular vesicles, named matrix vesicles (MVs), released by osteogenic cells. The MV membrane is enriched in sphingomyelin (SM), cholesterol (Chol) and tissue non-specific alkaline phosphatase (TNAP) compared with the parent cells' plasma membrane. TNAP is an ATP phosphohydrolase bound to cell and MV membranes via a glycosylphosphatidylinositol (GPI) anchor. Previous studies have shown that the lipid microenvironment influences the catalytic activity of enzymes incorporated into lipid bilayers. However, there is a lack of information about how the lipid microenvironment controls the ability of MV membrane-bound enzymes to induce mineral precipitation. Herein, we used TNAP-harboring proteoliposomes made of either pure dimyristoylphosphatidylcholine (DMPC) or DMPC mixed with either Chol, SM or both of them as MV biomimetic systems to evaluate how the composition modulates the lipid microenvironment and, in turn, TNAP incorporation into the lipid bilayer by means of calorimetry. These results were correlated with the proteoliposomes' catalytic activity and ability to induce the precipitation of amorphous calcium phosphate (ACP) in vitro. DMPC:SM proteoliposomes displayed the highest efficiency of mineral propagation, apparent affinity for ATP and substrate hydrolysis efficiency, which correlated with their highest degree of membrane organization (highest Delta H), among the tested proteoliposomes. Results obtained from turbidimetry and Fourier transformed infrared (FTIR) spectroscopy showed that the tested proteoliposomes induced ACP precipitation with the order DMPC:SM > DMPC:Chol:SM approximate to DMPC:Chol > DMPC which correlated with the lipid organization and the presence of SM in the proteoliposome membrane. Our study arises important insights regarding the physical properties and role of lipid organization in MV-mediated mineralization. (AU)

Processo FAPESP: 17/08892-9 - Superfícies bioativas obtidas a partir de filmes Langmuir-Blodgett e biominerais
Beneficiário:Ana Paula Ramos
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 14/00371-1 - Interações entre colágeno e proteínas/enzimas presentes nas vesículas da matriz são responsáveis pelo controle do processo de biomineralização?
Beneficiário:Maytê Bolean Correia
Linha de fomento: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 18/12092-0 - Estudos Biofísicos da Correlação Estrutura-função da Enzima TNAP em interação com modelos de membranas
Beneficiário:Bruno Zoccaratto Favarin
Linha de fomento: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 15/06814-5 - Interações entre colágeno e proteínas/enzimas presentes nas vesículas da matriz são responsáveis pelo controle do processo de biomineralização?
Beneficiário:Maytê Bolean Correia
Linha de fomento: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado