Improvement of two-dimensional gel electrophoresis proteome maps of the haloarchaeon Haloferax volcanii

Proteins of haloarchaea are remarkably unstable in low‐ionic‐strength solvents and tend to aggregate under standard two‐dimensional (2‐D) gel electrophoresis conditions, causing strong horizontal streaking. We have developed a new approach to generate 2‐D maps of halophilic proteins which included w...

Full description

Saved in:
Bibliographic Details
Published in:Proteomics (Weinheim) 2005-02, Vol.5 (2), p.354-359
Main Authors: Karadzic, Ivanka M., Maupin-Furlow, Julie A.
Format: Article
Language:English
Subjects:
Acetone - pharmacology
Analytical, structural and metabolic biochemistry
Archaea
Archaea - chemistry
Bacterial Proteins - chemistry
Bacterial Proteins - isolation & purification
Biological and medical sciences
Cholic Acids - pharmacology
Detergents - pharmacology
Electrophoresis, Gel, Two-Dimensional
Fundamental and applied biological sciences. Psychology
Haloarchaea
Haloferax volcanii
Haloferax volcanii - chemistry
Halophile
Hydrogen-Ion Concentration
Miscellaneous
Oxidation-Reduction
Peptide Mapping
Phosphines - pharmacology
Proteins
Proteome
Solubility
Thiourea - pharmacology
Two-dimensional gel electrophoresis
Urea - pharmacology
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Proteins of haloarchaea are remarkably unstable in low‐ionic‐strength solvents and tend to aggregate under standard two‐dimensional (2‐D) gel electrophoresis conditions, causing strong horizontal streaking. We have developed a new approach to generate 2‐D maps of halophilic proteins which included washing cells with 1.5 M Tris‐HCl buffer. In addition, proteins were precipitated with acetone, solubilized with urea and thiourea in the presence of the sulfobetaine detergent 3‐[(3‐cholamidopropyl)dimethylamino]‐1‐propanesulfonate (CHAPS), reduced with tributylphosphine (TBP), and separated with microrange strips of immobilized pH gradients (pH 3.9–5.1). This combination enabled the construction of highly reproducible 2‐D maps of Haloferax volcanii proteins.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200400950