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Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization

The biochemical machinery involved in matrix vesicles-mediated bone mineralization involves a specific set of lipids, enzymes, and proteins. Annexins, among their many functions, have been described as responsible for the formation and stabilization of the matrix vesicles′ nucleational core. However...

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Autores principales: Veschi, Ekeveliny Amabile, Bolean, Maytê, da Silva Andrilli, Luiz Henrique, Sebinelli, Heitor Gobbi, Strzelecka-Kiliszek, Agnieszka, Bandorowicz-Pikula, Joanna, Pikula, Slawomir, Granjon, Thierry, Mebarek, Saida, Magne, David, Millán, José Luis, Ramos, Ana Paula, Buchet, Rene, Bottini, Massimo, Ciancaglini, Pietro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9409191/
https://www.ncbi.nlm.nih.gov/pubmed/36012211
http://dx.doi.org/10.3390/ijms23168945
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author Veschi, Ekeveliny Amabile
Bolean, Maytê
da Silva Andrilli, Luiz Henrique
Sebinelli, Heitor Gobbi
Strzelecka-Kiliszek, Agnieszka
Bandorowicz-Pikula, Joanna
Pikula, Slawomir
Granjon, Thierry
Mebarek, Saida
Magne, David
Millán, José Luis
Ramos, Ana Paula
Buchet, Rene
Bottini, Massimo
Ciancaglini, Pietro
author_facet Veschi, Ekeveliny Amabile
Bolean, Maytê
da Silva Andrilli, Luiz Henrique
Sebinelli, Heitor Gobbi
Strzelecka-Kiliszek, Agnieszka
Bandorowicz-Pikula, Joanna
Pikula, Slawomir
Granjon, Thierry
Mebarek, Saida
Magne, David
Millán, José Luis
Ramos, Ana Paula
Buchet, Rene
Bottini, Massimo
Ciancaglini, Pietro
author_sort Veschi, Ekeveliny Amabile
collection PubMed
description The biochemical machinery involved in matrix vesicles-mediated bone mineralization involves a specific set of lipids, enzymes, and proteins. Annexins, among their many functions, have been described as responsible for the formation and stabilization of the matrix vesicles′ nucleational core. However, the specific role of each member of the annexin family, especially in the presence of type-I collagen, remains to be clarified. To address this issue, in vitro mineralization was carried out using AnxA6 (in solution or associated to the proteoliposomes) in the presence or in the absence of type-I collagen, incubated with either amorphous calcium phosphate (ACP) or a phosphatidylserine-calcium phosphate complex (PS–CPLX) as nucleators. Proteoliposomes were composed of 1,2-dipalmitoylphosphatidylcholine (DPPC), 1,2-dipalmitoylphosphatidylcholine: 1,2-dipalmitoylphosphatidylserine (DPPC:DPPS), and DPPC:Cholesterol:DPPS to mimic the outer and the inner leaflet of the matrix vesicles membrane as well as to investigate the effect of the membrane fluidity. Kinetic parameters of mineralization were calculated from time-dependent turbidity curves of free Annexin A6 (AnxA6) and AnxA6-containing proteoliposomes dispersed in synthetic cartilage lymph. The chemical composition of the minerals formed was investigated by Fourier transform infrared spectroscopy (FTIR). Free AnxA6 and AnxA6-proteoliposomes in the presence of ACP were not able to propagate mineralization; however, poorly crystalline calcium phosphates were formed in the presence of PS–CPLX, supporting the role of annexin-calcium-phosphatidylserine complex in the formation and stabilization of the matrix vesicles’ nucleational core. We found that AnxA6 lacks nucleation propagation capacity when incorporated into liposomes in the presence of PS–CPLX and type-I collagen. This suggests that AnxA6 may interact either with phospholipids, forming a nucleational core, or with type-I collagen, albeit less efficiently, to induce the nucleation process.
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spelling pubmed-94091912022-08-26 Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization Veschi, Ekeveliny Amabile Bolean, Maytê da Silva Andrilli, Luiz Henrique Sebinelli, Heitor Gobbi Strzelecka-Kiliszek, Agnieszka Bandorowicz-Pikula, Joanna Pikula, Slawomir Granjon, Thierry Mebarek, Saida Magne, David Millán, José Luis Ramos, Ana Paula Buchet, Rene Bottini, Massimo Ciancaglini, Pietro Int J Mol Sci Article The biochemical machinery involved in matrix vesicles-mediated bone mineralization involves a specific set of lipids, enzymes, and proteins. Annexins, among their many functions, have been described as responsible for the formation and stabilization of the matrix vesicles′ nucleational core. However, the specific role of each member of the annexin family, especially in the presence of type-I collagen, remains to be clarified. To address this issue, in vitro mineralization was carried out using AnxA6 (in solution or associated to the proteoliposomes) in the presence or in the absence of type-I collagen, incubated with either amorphous calcium phosphate (ACP) or a phosphatidylserine-calcium phosphate complex (PS–CPLX) as nucleators. Proteoliposomes were composed of 1,2-dipalmitoylphosphatidylcholine (DPPC), 1,2-dipalmitoylphosphatidylcholine: 1,2-dipalmitoylphosphatidylserine (DPPC:DPPS), and DPPC:Cholesterol:DPPS to mimic the outer and the inner leaflet of the matrix vesicles membrane as well as to investigate the effect of the membrane fluidity. Kinetic parameters of mineralization were calculated from time-dependent turbidity curves of free Annexin A6 (AnxA6) and AnxA6-containing proteoliposomes dispersed in synthetic cartilage lymph. The chemical composition of the minerals formed was investigated by Fourier transform infrared spectroscopy (FTIR). Free AnxA6 and AnxA6-proteoliposomes in the presence of ACP were not able to propagate mineralization; however, poorly crystalline calcium phosphates were formed in the presence of PS–CPLX, supporting the role of annexin-calcium-phosphatidylserine complex in the formation and stabilization of the matrix vesicles’ nucleational core. We found that AnxA6 lacks nucleation propagation capacity when incorporated into liposomes in the presence of PS–CPLX and type-I collagen. This suggests that AnxA6 may interact either with phospholipids, forming a nucleational core, or with type-I collagen, albeit less efficiently, to induce the nucleation process. MDPI 2022-08-11 /pmc/articles/PMC9409191/ /pubmed/36012211 http://dx.doi.org/10.3390/ijms23168945 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Veschi, Ekeveliny Amabile
Bolean, Maytê
da Silva Andrilli, Luiz Henrique
Sebinelli, Heitor Gobbi
Strzelecka-Kiliszek, Agnieszka
Bandorowicz-Pikula, Joanna
Pikula, Slawomir
Granjon, Thierry
Mebarek, Saida
Magne, David
Millán, José Luis
Ramos, Ana Paula
Buchet, Rene
Bottini, Massimo
Ciancaglini, Pietro
Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title_full Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title_fullStr Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title_full_unstemmed Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title_short Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization
title_sort mineralization profile of annexin a6-harbouring proteoliposomes: shedding light on the role of annexin a6 on matrix vesicle-mediated mineralization
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9409191/
https://www.ncbi.nlm.nih.gov/pubmed/36012211
http://dx.doi.org/10.3390/ijms23168945
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