Expression in Escherichia coli and purification of hexahistidine-tagged human tissue transglutaminase

Protein Expression and Purification
Qingli ShiArthur J L Cooper

Abstract

Recent evidence suggests that aberrant transglutaminase activity is associated with a wide variety of diseases. Tissue transglutaminase is the most widely distributed of the six well-characterized transglutaminases in humans. We describe a method for expressing hexahistidine-tagged human tissue transglutaminase in Escherichia coli BL21(DE3) using the pET-30 Ek/LIC expression vector. Purification of the expressed enzyme from suspensions of E. coli cells treated with CelLytic B Bacterial Cell Lysis/Extraction Reagent was accomplished by immobilized metal (Ni2+) affinity column chromatography. The procedure typically yields highly purified and highly active recombinant human tissue transglutaminase in about 1 day (about 0.6 mg/from a 1-liter culture).

References

Jan 12, 1978·Biochimica Et Biophysica Acta·S C Brenner, F Wold
Dec 1, 1972·Analytical Biochemistry·L LorandL Cooperstein
Nov 1, 1994·Bioscience, Biotechnology, and Biochemistry·H YasuedaM Motoki
Apr 7, 1997·Biochemical and Biophysical Research Communications·S Y KimJ Lee
Jun 23, 1999·Proceedings of the National Academy of Sciences of the United States of America·M V KarpujL Steinman
Apr 5, 2000·Progress in Neurobiology·M LesortG V Johnson
Apr 26, 2001·Preparative Biochemistry & Biotechnology·A Ambrus, L Fésüs
May 18, 2001·Analytical Biochemistry·T M JeitnerA J Cooper
Dec 12, 2001·Neurochemistry International·Bruce A CitronBarry W Festoff
Dec 12, 2001·Neurochemistry International·Soo Youl KimPeter M Steinert

❮ Previous
Next ❯

Citations

Oct 23, 2012·Protein Expression and Purification·Isabelle RoyJeffrey W Keillor
Dec 10, 2015·Frontiers in Plant Science·Vanesa S Marín ViegasSilvana Petruccelli
Sep 4, 2007·The Journal of Membrane Biology·Edgar Gutierrez, L Amy Sung
Jul 3, 2008·Amino Acids·Angela SorrentinoLoredana Mariniello
Aug 3, 2010·Expert Review of Vaccines·Linda AvesaniMario Pezzotti
Jan 1, 2013·Biomolecules·Natalie M Rachel, Joelle N Pelletier
Jan 8, 2004·Journal of Autoimmunity·Ken NakachiBernard Rees Smith
Jan 9, 2004·Protein Expression and Purification·Steve M F G GilletJoelle N Pelletier

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.