Deepak Kumar

Title: Insights of pseudoesterase activity of human serum albumin using 2-Naphthyl acetate and p-Nitrophenyl acetate: A LC-MS based approach

Abstract:

Human serum albumin (HSA) is the most abundant plasma protein. Various studies have shown that it also possesses pseudoesterase activity and the critical residue for such activity is Tyrosine 411. Estimation of HSA in serum and urine is important from the clinical point of view. Recently, it is shown that such enzymatic activity of HSA has the potential to regulate cholesterol biosynthesis and in detection of microalbuminuria. Therefore, understanding of pseudoesterase activity of HSA is critical. Several substrates are known for establishing the pseudoesterase activity of HSA. However, which ester substrate is the best substrate for HSA is not known. 
Objectives: To study the pseudoesterase activity of HSA with 2-Naphthyl acetate and p-Nitrophenyl acetate using LC-MS based approach.
Methods: In-silico studies (Molecular docking and Molecular Dynamics simulations studies) and in-vitro studies were carried out.
Results: Acetylation of HSA at various residues was observed in both cases but with a different pattern.
Conclusion: Based on the data we believe that 2-Naphthyl acetate-induced acetylation is readily detectable by LC-MS and the substrate overcomes the limitations of p-nitrophenylacetate-based pseudoesterase activity determination

Audience Take Away Notes :

• The audience will learn about HSA (most abundant plasma protein) and its pseudoesterase activity. Such activity hass the potential to regulate various metabolic processes and pharmacokinetics of various ester drugs like aspirin. 
• The talk will discuss the analysis of various in-silico and experimental data. Thus, the audience can learn the methodology and analysis aspect of the same. 
• The physiological significance of pseudoesterase activity of HSA is not known yet. It is believed that pseudoesterase activity of HSA has the potential to regulate cholesterol biosynthesis. Therefore, the talk will broaden the horizon of understanding/tackling metabolic disorders like atherosclerosis, NAFLD, etc.
• Understanding the pseudeosterase activity of HSA which will pave path for designing a point of care method for microalbuminuria detection.
• The pseudeosterase activity-based detection of albumin has a potential to overcome the limitations of dye-based and immunochemical based methods for microalbuminuria detection

Biography:

Deepak Kumar has done his PhD degree working under the supervision of Prof Dibyajyoti Banerjee in 2021 from PGIMER, Chandigarh, India. The title of his PhD work is “Development of a novel strategy for rapid detection of human serum albumin pseudoesterase activity – A step towards a novel point of care screening for microalbuminuria”. He joined as Junior Demonstrator in the Department of Experimental Medicine and Biotechnology, PGIMER, Chandigarh in 2021 and teaching postgraduate students. In 2022 he joined as Senior Demonstrator in the same Department and continuing teaching and his research work. He is working to understand the effect of post-translational modification of the enzymatic activity of HSA and other relevant enzymes. Moreover, with his team he is studying the effect of bisphenols on E. coli and HepG2 cells. He has more than 25 publications which consist of research articles, book chapters, etc. in reputed journals and publishing house.