02197nas a2200217   4500008004100000022001400041245017500055210006900230260007900299300001200378490000900390520129600399653001501695653002401710653001901734653003601753653003201789100002101821700001401842856012301856       2007                        eng d  a0096-300300aBiochemical characterization of an aspartic protease from vigna radiata: kinetic interactions with the classical inhibitor pepstatin implicating a tight binding mechanism0 aBiochemical characterization of an aspartic protease from vigna   a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcMAY  a619-6270 v17743 a<p>{Aspartic proteases are the focus of recent research interest in understanding the physiological importance of this class of enzymes in plants. This is the first report of an aspartic protease from the seeds of Vigna radiata. The aspartic protease was purified to homogeneity by fractional ammonium sulfate precipitation and pepstatin-A agarose affinity column. It was found to have a molecular weight of 67,406 Da by gel filtration chromatography. SDS-PAGE analysis revealed the presence of a heterodimer with subunits of molecular weights of 44,024 and 23,349 Da respectively. The enzyme was pH stable with the amino acid analysis confirming the molecular weight of the protein. The substrate cleavage site as analyzed by using the synthetic substrate was found to be the Phe-Tyr bond. The kinetic interactions of the enzyme were studied with the universal inhibitor, pepstatin A. This is the first report on the interactions of a plant aspartic protease with pepstatin-A, an inhibitor from a microbial source. A competitive one-step mechanism of binding is observed. The progress curves are time-dependent and consistent with tight binding inhibition. The K(i) value of the reversible complex of pepstatin with the enzyme was 0.87 mu M whereas the overall inhibition constant K(i)*</p>10aant colony10aglobal optimization10ametaheuristics10amultimodal continuous functions10aparticle swarm optimization1 aKulkarni, Aarohi1 aRao, Mala  uhttps://library.ncl.res.in/content/biochemical-characterization-aspartic-protease-vigna-radiata-kinetic-interactions-0