Isolate, Purify And Characterise Metallothionein From The Hepatic Cells Of Oreochromis Niloticus [ Linnaeus, 1758]
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Abstract
Objective(s): The present study intends to isolate and purify Metallothionein extracted from the liver tissue of Oreochromis niloticus exposed to Cadmium. Study of MT will help to know its protective role against Cadmium
Method(s): Acclimatised fishes were treated with sub - lethal concentration of Cadmium to induce the synthesis of Metallothionein. MT was isolated and purified from fish liver by affinity chromatography from differential distribution (3KD – 43KD) across eluted fractions. Molecular weight of the purified elute was analysed by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) and Peptide Mass Fingerprinting and confirmed to be Metallothionein.
Findings: Different elute fractions were obtained from the MT specific Affinity Column chromatography. At each fraction-by-fraction purification led to obtaining a single band in the seventh elute fraction. This single band was analysed using MALDI-TOF. Trypsin digestion produced a fragmented peptide mass spectrum. The mass spectrogram revealed a single peak, indicating purified MT protein with a molecular weight of 6140.52 Daltons (~6.2 kDa). The finding contributes valuable insights into Metallothionein structure and properties, and understanding its role in metal detoxification and stress regulating cellular processes.
Novelty: Molecular weight of the Metallothionein from liver tissue of Oreochromis niloticus was found using MALDI-TOF. This method is a simple way to isolate and purify low molecular weight protein like MT.
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