Analysis of cellular phosphoproteins by two‐dimensional gel electrophoresis: Applications for cell signaling in normal and cancer cells

Guy, Graeme R.; Philip, R; Tan, Y. H.

doi:10.1002/elps.1150150160

Cited by 82 publications

(62 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Yeast Dbf2p kinase and V-ATPase function indicate differential levels of phosphorylation (Guy et al, 1994). The absence of a more acidic Vma2p isoform in dbf2D phosphoprotein extracts is therefore consistent with loss of a more phosphorylated Vma2p isoform.…”

Section: Loss Of Dbf2p Is Associated With Reduced Phosphorylation Of mentioning

confidence: 68%

A novel role for the yeast protein kinase Dbf2p in vacuolar H+-ATPase function and sorbic acid stress tolerance

et al. 2007

View full text Add to dashboard Cite

In Saccharomyces cerevisiae, the serine-threonine protein kinase activity of Dbf2p is required for tolerance to the weak organic acid sorbic acid. Here we show that Dbf2p is required for normal phosphorylation of the vacuolar H + -ATPase (V-ATPase) A and B subunits Vma1p and Vma2p.Loss of V-ATPase activity due to bafilomycin treatment or deletion of either VMA1 or VMA2 resulted in sorbic acid hypersensitivity and impaired vacuolar acidification, phenotypes also observed in both a kinase-inactive dbf2 mutant and cells completely lacking DBF2 (dbf2D). Crucially, VMA2 is a multicopy suppressor of both the sorbic acid-sensitive phenotype and the impaired vacuolar-acidification defect of dbf2D cells, confirming a functional interaction between Dbf2p and Vma2p. The yeast V-ATPase is therefore involved in mediating sorbic acid stress tolerance, and we have shown a novel and unexpected role for the cell cycle-regulated protein kinase Dbf2p in promoting V-ATPase function.

show abstract

Section: Loss Of Dbf2p Is Associated With Reduced Phosphorylation Of mentioning

confidence: 68%

A novel role for the yeast protein kinase Dbf2p in vacuolar H+-ATPase function and sorbic acid stress tolerance

et al. 2007

View full text Add to dashboard Cite

show abstract

“…There are various methods that can be used to concentrate diluted protein samples for electrophoresis, including ultrafiltration, 6,12) and precipitation with acetone, 6,8) trichloroacetic acid (TCA), 8,11) and TCA/ acetone. 6,9,10) These methods can also be used for urine.…”

Section: Resultsmentioning

confidence: 99%

“…For acetone precipitation, urine was added to four times the volume of cold acetone as described elsewhere, with modifications. 6,8) The mixture was stored for 3 hr at -30°C, after which it was centrifuged at 10000 g at 4°C for 15 min. The pellet thus obtained was washed with a cold diethylether : ethanol (6 : 1) mixture and centrifuged at 10000 g at 4°C for 5 min.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Improved Recovery of Human Urinary Protein for Electrophoresis

Fujimoto

Miya

Machida

et al. 2006

JOURNAL OF HEALTH SCIENCE

View full text Add to dashboard Cite

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)is frequently used to analyze urinary proteins, but appropriate urine sample preparation is essential, because urinary proteins are present in small concentrations, and urine contains high concentrations of salts and metabolic wastes. The aim of this study was to determine the optimum method for urine sample preparation. The effects on urinary protein recovery of pretreating the ultrafiltration apparatus with various agents (Triton X-100, Tween20, PEG compound, and SDS) were studied; Triton X-100 was found to be the most effective agent. Ultrafiltration and acetone precipitation were compared as sample preparation methods for SDS-PAGE. Recovery of urinary protein using ultrafiltration (89.5 ± 8.1%) was better than that achieved by using acetone precipitation (75.1 ± 12.2%) (p < 0.01). Integrated densitometric values for five protein bands (62, 52, 39, 31, and 6 kDa) were higher for ultrafiltration samples than for acetone samples (42350 ± 2568 vs. 37010 ± 725, 34665 ± 1519 vs. 32355 ± 2278, 10755 ± 1683 vs. 8870 ± 941, 65920 ± 3354 vs. 60680 ± 3651, and 1355 ± 160 vs. 1130 ± 211, respectively) (p < 0.05), although differences for the other nine bands were not significant. In conclusion, ultrafiltration is a suitable method for preparing urine samples for SDS-PAGE.

show abstract

“…32 P]phosphate, followed by 2-DE [36,37]. Phosphorylations on serine, threonine and tyrosine residues have been studied using b-elimination-directed cleavage at the site of modification, by fluorescent dyes, and by chemical modification of phosphorylated residues [20][21][22]38].…”

Section: Functional Status Of Proteinsmentioning

confidence: 99%

Bridging proteomics and systems biology: What are the roads to be traveled?

Souchelnytskyi

2005

Proteomics

View full text Add to dashboard Cite

The comprehensive study of proteomes has become an important part of attempts to uncover the systemic properties of biological systems. Proteomics provides data of a quality which increasingly fulfills strict requirements of systems biology for quantitative and qualitative information. Notably, proteomics can generate rich datasets that describe dynamic changes of proteomes. On the other hand, large-scale modeling requires the development of mathematic tools that are adequate for the processing of largely uncertain biological data. In this review, recent developments that pave the way for the integration of proteomics into systems biology are discussed. These developments include the standardization of data acquisition and presentation, the increased comprehensiveness of proteomics studies in description of functional status, localization and dynamics of proteins, and advanced modeling approaches. Proteomics and modeling of biological processesThe dynamic complexity of biological processes has been less well understood, when compared to many physical and chemical processes. Apparently, sending a man to the Moon is less complicated than the full understanding of how bruises heal. This situation is about to change: the sequencing of a number of genomes, large-scale explorations of transcriptomes, proteomes and metabolomes, and a huge volume of directed studies inspire hope that we will be able to describe a living creature in the strict language of mathematics. Most importantly, there is a hope that we will be able to design better treatments and predict outcomes for human diseases. The development of modeling tools fuels these expectations, with the dawn of systems biology. Study of the systemic properties of biological systems, as systems biology can be defined, has already provided successful examples, e.g., insights into the physiology of the heart, diabetes, asthma and cancer (reviewed in [1,2]).Building and analysis of models of biological processes comprises a number of modeling tools, and addresses biological complexity on the levels from biochemical reactions and cell physiology to behavior and evolution [3,4]. Here and through-out this review the term "model" refers to description of biological processes in mathematical terms, without discrimination of mathematical tools. Consequently, modeling is defined as "the application of methods to analyze complex, real-world problems to make predictions about what might happen with various actions" (see Computational Science Glossary, wofford.info/ecs/glossary/ terms.htm; Table 1). Modeling tools cover a broad range of mathematical methods, from systems of differential equations to statistical correlation tools [3,4]. Some of the tools require detailed knowledge about components, e.g., to build a systems of differential equations for modeling of a signaling Abbreviations: ABPP, activity-based proteome profiling; BEMAD, b-elimination followed by Michael addition with DTT; FAK, focal adhesion kinase; FRET, fluorescence resonance energy transfer; GFP, green fluores...

show abstract

Analysis of cellular phosphoproteins by two‐dimensional gel electrophoresis: Applications for cell signaling in normal and cancer cells

Cited by 82 publications

References 70 publications

A novel role for the yeast protein kinase Dbf2p in vacuolar H+-ATPase function and sorbic acid stress tolerance

A novel role for the yeast protein kinase Dbf2p in vacuolar H+-ATPase function and sorbic acid stress tolerance

Improved Recovery of Human Urinary Protein for Electrophoresis

Bridging proteomics and systems biology: What are the roads to be traveled?

Contact Info

Product

Resources

About