Journal ArticleDOI
Enrichment of phosphorylated proteins and peptides from complex mixtures using metal oxide/hydroxide affinity chromatography (MOAC).
TLDR
A novel method termed metal oxide affinity chromatography (MOAC) of enriching for phosphorylated proteins and peptides based on the affinity of the phosphate group for Al(OH)3 is presented here and is more selective, more cost effective and easily applicable to method optimization.Abstract:
A novel method termed metal oxide affinity chromatography (MOAC) of enriching for phosphorylated proteins and peptides based on the affinity of the phosphate group for Al(OH)(3) is presented here. When compared to commercial phosphoprotein-enrichment kits, this method is more selective, more cost effective and easily applicable to method optimization. The use of glutamic and aspartic acid in the loading buffer significantly enhances selectivity. Standard protein mixtures and complex Arabidopsis thaliana leaf protein extracts were tested for efficacy of enrichment. The method can be applied to proteins extracted using either mild or denaturing conditions. The same Al(OH)(3) material is suitable for the enrichment of phosphopeptides out of a tryptic digest of alpha-casein. Peptide phosphorylation was revealed by beta-elimination of phosphate groups. Enrichment and in vivo phosphorylation of A. thaliana leaf proteins were confirmed with Pro-Q diamond stain. Several of the phosphoprotein candidates that were identified by MS are known to be phosphorylated in vivo in other plant species.read more
Citations
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Journal ArticleDOI
Selective zirconium dioxide-based enrichment of phosphorylated peptides for mass spectrometric analysis.
TL;DR: This work presents the first demonstration of the utility of zirconium dioxide microtips for phosphopeptide isolation prior to mass spectrometric analysis, and shows that these microtips display similar overall performance as TiO2 microtips.
Journal ArticleDOI
Phosphopeptide Enrichment by Aliphatic Hydroxy Acid-modified Metal Oxide Chromatography for Nano-LC-MS/MS in Proteomics Applications
Naoyuki Sugiyama,Takeshi Masuda,Kosaku Shinoda,Akihiro Nakamura,Masaru Tomita,Yasushi Ishihama,Yasushi Ishihama +6 more
TL;DR: This is the first successful application of a single MOC-based approach to phosphopeptide enrichment from complex biological samples such as cell lysates, and the combined results provided 1646 uniqueosphopeptides.
Journal ArticleDOI
Modification-specific proteomics: strategies for characterization of post-translational modifications using enrichment techniques
Yingming Zhao,Ole N. Jensen +1 more
TL;DR: Modification‐specific enrichment techniques combined with advanced MS/MS methods and computational data analysis have revealed a surprisingly large extent of PTMs in proteins, including multi‐site, cooperative modifications in individual proteins.
Journal ArticleDOI
Quantitative Proteomic Analysis of Histone Modifications
TL;DR: Dysregulation of these processes has been intimately associated with the development of diseases such as cancer and is thought to regulate chromatin structure and function by two mechanisms.
Journal ArticleDOI
Development of deep eutectic solvents applied in extraction and separation.
Xiaoxia Li,Kyung Ho Row +1 more
TL;DR: The available data and references in this field are reviewed to summarize the applications and developments of deep eutectic solvents.
References
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Journal ArticleDOI
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal ArticleDOI
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Journal Article
Cleavage of structural proteins during the assemble of the head of bacterio-phage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Journal ArticleDOI
A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids
D. Wessel,U.I. Flügge +1 more
TL;DR: A rapid method based on a defined methanol-chloroform-water mixture for the quantitative precipitation of soluble as well as hydrophobic proteins from dilute solutions (e.g., column chromatography effluents) has been developed.
Journal ArticleDOI
Tandem Mass Tags: A Novel Quantification Strategy for Comparative Analysis of Complex Protein Mixtures by MS/MS
Andrew J. Thompson,Jürgen Schäfer,Karsten Kuhn,Stefan Kienle,Josef Schwarz,Günter Schmidt,and Thomas Neumann,Christian Hamon +7 more
TL;DR: A novel MS/MS-based analysis strategy using isotopomer labels, referred to as "tandem mass tags" (TMTs), for the accurate quantification of peptides and proteins is described, which allows peptides from different samples to be identified by their relative abundance with greater ease and accuracy than other methods.