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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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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]
Total Authors: 9
[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
Total Affiliations: 5
Document type: Journal article
Source: Archives of Biochemistry and Biophysics; v. 691, SEP 30 2020.
Web of Science Citations: 0

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)

FAPESP's process: 17/08892-9 - Bioactive surfaces designed from Langmuir-Blodgett Films and Biominerals
Grantee:Ana Paula Ramos
Support Opportunities: Regular Research Grants
FAPESP's process: 14/00371-1 - Are the interactions between collagen and proteins/enzymes present in the matriz vesicles responsible for the control in the biomineralization process?
Grantee:Maytê Bolean Correia
Support Opportunities: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 18/12092-0 - Biophysical Studies of the Structure-function Correlation of TNAP in interaction with membrane models
Grantee:Bruno Zoccaratto Favarin
Support Opportunities: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 15/06814-5 - Do interactions between collagen and proteins/enzymes present in matrix vesicles control biomineralization?
Grantee:Maytê Bolean Correia
Support Opportunities: Scholarships abroad - Research Internship - Post-doctor