A unified method for purification of basic proteins.

Analytical Biochemistry
Sanjay AdhikariRabindra Roy

Abstract

Protein purification is still very empirical, and a unified method for purifying proteins without an affinity tag is not available yet. In the postgenomic era, functional genomics, however, strongly demands such a method. In this paper we have formulated a unique method that can be applied for purifying any recombinant basic protein from Escherichia coli. Here, we have found that if the pH of the buffer is merely one pH unit below the isoelectric point (pI) of the recombinant proteins, most of the latter bind to the column. This result supports the Henderson-Hasselbalch principle. Considering that E. coli proteins are mostly acidic, and based on the pI determined theoretically, apparently all recombinant basic proteins (at least pI-1 > or = 6.94) may be purified from E. coli in a single step using a cation-exchanger resin, SP-Sepharose, and a selected buffer pH, depending on the pI of the recombinant protein. Approximately, two-fifths of human proteome, including many if not all nucleic acid-interacting proteins, have a pI of 7.94 or higher; virtually all these 12,000 proteins may be purified using this method in a single step.

References

Dec 26, 2001·Nucleic Acids Research·T HubbardM Clamp
Apr 2, 2005·Protein Expression and Purification·Jordan J LichtySong Tan
Feb 24, 2006·Nature·Laura Bonetta
Aug 12, 2006·The Journal of Biological Chemistry·Sanjay AdhikariRabindra Roy
Aug 25, 2007·The Journal of Biological Chemistry·Sanjay AdhikariRabindra Roy
Jun 6, 2009·Analytical Biochemistry·Ming-Kai ChernChien-Wei Liu

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Citations

Mar 26, 2011·Journal of Molecular Evolution·Varinia López-RamírezGabriela Olmedo-Álvarez
Nov 2, 2011·Molecular and Cellular Biochemistry·Sanjay AdhikariRabindra Roy
Jan 9, 2017·The FEBS Journal·Yuniel Fernández-MarreroAna J Garcia-Saez
Feb 9, 2019·Journal, Genetic Engineering & Biotechnology·Sadam D V SatyanarayanaSirisha Jeereddy
Oct 8, 2020·Biomedical Chromatography : BMC·Edouard C Nice

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