Among various enzyme families’ proteases are considered as a very important group as it plays very important role in various biological processes. Any malfunctioning in proteases can cause various disorders like cancer, infections and deteriorating disorders.
Due to this importance of proteases several inquiries are made for studying about proteases and this has led to development of various methods for analyzing substrate specificity. In the entire process it requires analyzing substrate specificity of a huge number of proteases and target molecules, therefore there is requirement of high throughput screening methods. This has influenced Beata Bozoki et al to develop a method for analyzing proteases that is HTS compatible by using recombinant fusion protein as substrates. It was published in January 2018 in Analytical Biochemistry. This protein that is used as substrate consist of many N terminal, fusion labels, cleavage classification that is similar to tobacco etch virus, HIV-1 proteases and C-terminal fluorescent protein.
The methodology mainly involves fluorescents proteins are detected fluorometrically that are actually released by proteolytic cleavage from the magnetic bead attached substrates. This method will help the researchers in activity quantities of proteases like TEV and HIV-1 which in turn can be used for testing the appropriateness of the system for enzyme kinetic dimensions, hang-up studies and determining pH optimum.
