A Proteomic Approach for The Study of Macrotyloma Uniflorum (Horsegram) Seed Proteins for Kidney Stone Degradation: Conceptual Study in Cultivars of Himachal Pradesh, India

Proteomics is one of the complex and significant methodologies to know the genetic functions of proteins.The study of plant proteins having antilithiatic activity is an unexplored area and can be hit through proteomics.The present study was framed for in-depth identification and characterization of antilithiatic proteins from horse gram (Macrotyloma uniflorum).Total protein of Macrotyloma uniflorum seeds from six different location of Himachal Pradesh, India was isolated in protease inhibitor buffer.The identification of cultivar seed proteins was done by Matrix-assisted laser desorption/ionization-time of flight Mass spectrometry (MALDI-TOF MS).The results were framed by database search with the MASCOT server.The protein sample of Rampur location contained maximum number of peptides followed by Sundernagar, Palampur and Kullu.Seeds of Chamba location showed the presence of very few peptides.The quantification of Macrotyloma uniflorum seed proteins was done with Bradford method and further protein with high oxalate inhibition activity (15µg) was loaded on 12% Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gel.In the proteomic study of loaded protein, a total of three major low molecular weight protein bands were observed (~27KDa, ~23.1 and ~17KDa respectively).Seed protein SDS-PAGE may be used to investigate genetic diversity and relatedness in cultivars at a low cost.Furthermore, the proteomic analysis of the seed protein of M. uniflorum aids in the creation of a proteinbased marker that can be used to identify cultivars in the future.