Differential effects of the lipidic and ionic microenvironment on NPP1's phosphohydrolase and phosphodiesterase activities

Author(s)

Andrilli, Luiz H. S. ; Sebinelli, Heitor G. ; Cominal, Jucara G. ; Bolean, Mayte ; Hayann, Larwsk ; Millan, Jose Luis ; Ramos, Ana P. ; Ciancaglini, Pietro

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES; v. 1866, n. 4, p. 13-pg., 2024-02-13.

Abstract

Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) is an enzyme present in matrix vesicles (MV). NPP1 participates on the regulation of bone formation by producing pyrophosphate (PPi) from adenosine triphosphate (ATP). Here, we have used liposomes bearing dipalmitoylphosphatidylcholine (DPPC), sphingomyelin (SM), and cholesterol (Chol) harboring NPP1 to mimic the composition of MV lipid rafts to investigate ionic and lipidic influence on NPP1 activity and mineral propagation. Atomic force microscopy (AFM) revealed that DPPC-liposomes had spherical and smooth surface. The presence of SM and Chol elicited rough and smooth surface, respectively. NPP1 insertion produced protrusions in all the liposome surface. Maximum phosphodiesterase activity emerged at 0.082 M ionic strength, whereas maximum phosphomonohydrolase activity arose at low ionic strength. Phosphoserine-Calcium Phosphate Complex (PS-CPLX) and amorphous calcium -phosphate (ACP) induced mineral propagation in DPPC- and DPPC:SM-liposomes and in DPPC:Chol-liposomes, respectively. Mineral characterization revealed the presence of bands assigned to HAp in the mineral propagated by NPP1 harbored in DPPC-liposomes without nucleators or in DPPC:Chol-liposomes with ACP nucleators. These data show that studying how the ionic and lipidic environment affects NPP1 properties is important, especially for HAp obtained under controlled conditions in vitro. (AU)

• FAPESP's process: 21/03697-9 - Spectrophotometer with UV-RAMAN Optical Fiber - tematic project 2019/08568-2
• Grantee: Pietro Ciancaglini
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 22/04885-6 - Investigation of the role of Extracellular Vesicles (EVs) and Matrix Vesicles (MVs) in the initiation, propagation, regeneration and control of biological mineralization
• Grantee: Juçara Gastaldi Cominal
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 21/13140-1 - Study of PHOSPHO1 and nSMase2 interaction with model membranes: a possible correlation in matrix vesicle secretion
• Grantee: Luiz Henrique da Silva Andrilli
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 19/25054-2 - Strontium-containing nanoparticles and their versatility for biomaterials fabrication: implications and applications in biomineralization
• Grantee: Ana Paula Ramos
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/08568-2 - Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization
• Grantee: Pietro Ciancaglini
• Support Opportunities: Research Projects - Thematic Grants