In Silico Analysis of Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF)Gene to Increase the Stability of Gene Product- A Bioinformatics Approach

摘要

Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) is one of theseveral naturally occurring glycoproteins that regulate leukocyte production,migration and function. It has been produced in different cell types and withdifferent properties based on the production process. The objective of thiswork is to carry out the site-directed mutagenesis (in-silico analysis) in humanGranulocyte Macrophage Colony Stimulating Factor in order to increase thestability and half-life of this protein without affecting its biological activity.Among the other GM-CSF's, Murine GM-CSF is the most stable form. Hencein this work, sequential analysis of the murine and human GM-CSF wasperformed in detail to understand the differences in the stability. It was foundthat the sialic acid content increased the half life of GM-CSF. Based onphylogenetic analysis and the information about the interaction of sialic acid,number of point mutations with Glutamic Acid (Glu), Asparagine (Asn),Arginine (Arg) were implemented at position 78 (Threonine-Thr) of hGM-CSF and studied the effect of these mutations using Bio-Informatics tools.Energy minimisation studies were carried out to identify the stability. Thesestudies revealed the potential advantages of using Arg to increase the stability.Further, using Bio-Informatics tools such as PatchDock, Argus Lab and Hex.