Investigation of structural and functional properties of mnemiopsin۲ mutant (F۱۹۰L) after immobilization on silica

Background and Aim: Following the development of biotechnology, there is an increasing interest in the use of safe and sensitive tools related to biological phenomena such as fluorescence, chemoluminescence and bioluminescence. In temperament, there are two types of bioluminescence systems, including the luciferin-luciferase and the photoprotein. Due to their unique properties, photoproteins are used in many intracellular and extracellular studies, such as gene expression analysis, the pharmaceutical industry, biochemical processes, and imaging of living cells. The objective of this study was to investigate the changes in the emission and stability properties of a silica nanoparticles-immobilized mutant (F۱۹۰L) of mnemiopsin۲, as a calcium-regulating photoprotein, and to compare them with non-immobilized photoprotein under different assay conditions.Methods: . For this, white silica nanoparticles were firstly synthesized, and then the pure photoprotein was fixed on their surface using proprietary EDC / NHS linkers. All the conditions and parameters needed for the activity of photoprotein such as temperature, pH, amount of calcium, and half-life were monitored after stabilization, and they were compared with the optimized conditions for free photoprotein.Results: Results indicated that the silica nanoparticles-immobilized photoprotein had more stability and tendency to substrate, as well as higher optimal pH, while lower Ca۲+ sensitivity as compared to free photoprotein.Conclusion: According to the research, photoprotein has become more sensitive to pH after stabilization on silica nanoparticle and the relative temperature stability, half-life, calcium sensitivity and and tendency to substrate have increased, each of which can be used as a unique feature for different applications.

کلیدواژه ها:

Photoprotein ، Mnemiopsin۲ ، Immobilization ، Silica nanoparticle.