Enhanced Soluble Production of Biologically Active Recombinant Human p38 Mitogen-Activated-Protein Kinase (MAPK) in Escherichia coli
Authors: Sunil K. Khattar; Pankaj Gulati; Prabuddha K. Kundu; Vibhuti Singh; Usha Bughani; Malini Bajpai; Kulvinder. S. Saini
Source: Protein and Peptide Letters, Volume 14, Number 8, August 2007 , pp. 756-760(5)
Publisher: Bentham Science Publishers
Abstract:The conditions were optimized for maximum soluble yield of biologically active recombinant p38α mitogen activated protein kinase (MAPK) vis-a-vis insoluble fraction (inclusion body formation). This study reports a rapid, economical and single step purification process for the overproduction of GST tagged p38α MAPK. A yield of 18 mg of highly purified and soluble protein per liter of bacterial culture within 6 h timeframe was achieved. The purified protein was found to be biologically suitable for phosphorylation by upstream kinases and was catalytically active. We further demonstrated that our in-house p38α MAPK is more potent (>30%) than a commercially available enzyme.
Document Type: Research Article
Publication date: 2007-08-01
- Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.