A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies.

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Abstract Text:

Shiluan Yi,Anne Brickenden,Wing-Yiu Choy,Shiluan Yi,Anne Brickenden,Wing-Yiu Choy,

Human prothymosin alpha (ProTalpha) is a small acidic protein (12.1kDa; pI approximately 3.5) ubiquitously expressed in a wide variety of tissues. The amino acid composition of this protein is highly unusual. While close to half of its sequence is composed of acidic amino acids, the protein does not contain any aromatic residues. ProTalpha has been shown to play crucial roles in different biological processes including cell proliferation, transcriptional regulation and apoptosis. Despite the multiple functions this protein has, it does not adopt a stable tertiary fold under physiological conditions. In order to understand how ProTalpha functions, detailed structural characterization of this protein is essential. Nuclear magnetic resonance (NMR) spectroscopy is a powerful technique for elucidating the protein structure and dynamics at the atomic level. However, milligrams of isotopically labeled protein with high purity are usually required for the studies. In this work, we developed a high-yield protocol for purifying recombinant ProTalpha expressed in Escherichia coli by exploiting the intrinsically disordered and acidic natures of this protein. By combining the heat-cooling extraction, ammonium sulfate precipitation, and anion exchange chromatography, we were able to obtain over 20mg of ProTalpha with >97% purity from 1L of M9 minimal media culture. The new purification protocol provides a cost effective and an efficient way to produce large quantities of high purity recombinant human ProTalpha in various isotopically labeled forms, which will greatly facilitate the structural studies of this protein by NMR and other biophysical methods.

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Publishing Authors By Initials

S Yi,A Brickenden,WY Choy,S Yi,A Brickenden,WY Choy,

For similar abstracts research abstracts see: abstracts research

PUBMED ID PMID:

MEDLINE DATE:

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Protein expression and purification

VOLUME: 57

Page Numbers: 1-8

Journal Abbreviation: Protein Expr. Purif.

ISSN: 1046-5928

DAY: 14

MONTH: 09

YEAR: 2007

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9101496

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Keywords Mesh Terms:

KEYWORDS:

MESH TERMS:

Chemical & Substance for Abstract: A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies. Information

Substance Name:

Registry Number:

Grant and Affiliation Information for A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies.

AFFILIATION: Department of Biochemistry, The University of Western Ontario, London, Ont., Canada N6A 5C1.

Country: United States

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: Protein Expr Purif

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies Related Publications

• 'Stockbroker's knee'.
• 15N NMR spin relaxation dispersion study of the molecular crowding effects on protein folding under native conditions.
• A new protocol for high-yield purification of recombinant human prothymosin alpha expressed in Escherichia coli for NMR studies.
• A new repository penicillin (a form of aqueous penicillin G procaine) in infants and children.
• A phase I trial of outpatient weekly irinotecan/carboplatin and concurrent radiation for stage III unresectable non small-cell lung cancer: a Vanderbilt-Ingram Cancer Center Affiliate Network Trial.
• Anticancer drug-inorganic nanohybrid and its cellular interaction.
• Cellular toxicity of inorganic hydroxide nanoparticles.
• Changes in activity level in women aged 40-80 years.
• Differential effects of dietary vitamin E and antioxidants on eicosanoid synthesis in young rabbits.
• Dry eye and blink rate simulation with a pig eye model.
• EULAR evidence-based recommendations for the management of fibromyalgia syndrome.
• Effect of copper doping on the crystal structure and morphology of 1D nanostructured manganese oxides.
• Enhanced contrast of electrochromic full cell systems with nanocrystalline PEDOT-prussian blue.
• Hidden charge 2e Boson in doped Mott insulators.
• INDUCTION OF INCREASED FIBRINOLYTIC ACTIVATOR IN PLASMA BY VARIOUS DRUGS.
• Joint inflammation and cytokine inhibition in rheumatoid arthritis.
• Keratoacanthoma and infundibulocystic squamous cell carcinoma.
• Lead zirconate titanate/poly(vinylidene fluoride-trifluoroethylene) 1-3 composites for ultrasonic transducer applications.
• Mixed micelle-template route to mesoporous silica.
• Noninvasive Real-time Imaging of Tumors and Metastases Using Tumor-targeting Light-emitting Escherichia coli.
• Piezoelectric and pyroelectric properties of PZT/P(VDF-TrFE) composites with constituent phases poled in parallel or antiparallel directions.
• Point injection as an alternative acupuncture technique - an exploratory study of responses in healthy subjects.
• Potential for combined modality therapy of cyclooxygenase inhibitors and radiation.
• Sciatic pain.
• Soft-chemical synthesis and electrochemical characterization of multicomponent Mn(1-x-y)Co(x)Ni(y)O2 nanostructures.
• Synthesis of new visible light active photocatalysts of Ba(In(1/3)Pb(1/3)M'(1/3))O3 (M' = Nb, Ta): a band gap engineering strategy based on electronegativity of a metal component.
• Test characteristics of a level I or II fetal ultrasound in detecting structural heart disease.
• The benefits of in-home pharmacy evaluation for older persons.
• The intrinsically disordered TC-1 interacts with Chibby via regions with high helical propensity.
• Time-dependent X-ray absorption spectroscopic (XAS) study on the transformation of zinc basic salt into bis(N-oxopyridine-2-thionato) zinc (II).