Native‐DIGE: A new look at the mitochondrial membrane proteome

Dani, Diksha; Dencher, Norbert A.

doi:10.1002/biot.200800030

Cited by 19 publications

(11 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Pre‐labeling of protein samples by reactive fluorescent dyes or fusion with fluorescent proteins before application to native gels has been used only a few times so far 5, 25–31. Since Coomassie‐dye quenches fluorescence by around 95% 5, 1‐D BN gels are not immediately suitable for fluorescence detection.…”

Section: Development Of Techniques Associated With Bne and Cnementioning

confidence: 99%

Native electrophoretic techniques to identify protein–protein interactions

Wittig

Schägger

2009

Proteomics

View full text Add to dashboard Cite

Permanent protein-protein interactions are commonly identified by co-purification of two or more protein components using techniques like co-immunoprecipitation, tandem affinity purification and native electrophoresis. Here we focus on blue-native electrophoresis, clear-native electrophoresis, high-resolution clear-native electrophoresis and associated techniques to identify stable membrane protein complexes and detergent-labile physiological supercomplexes. Hints for dynamic protein-protein interactions can be obtained using two-hybrid techniques but not from native electrophoresis and other protein isolation techniques except after covalent cross-linking of interacting proteins in vivo prior to protein separation.

show abstract

Section: Development Of Techniques Associated With Bne and Cnementioning

confidence: 99%

Native electrophoretic techniques to identify protein–protein interactions

Wittig

Schägger

2009

Proteomics

View full text Add to dashboard Cite

show abstract

“…The advantage of DIGE can be extended to study membrane proteins and protein-protein interactions, by using it in combination with other gel based methods (Perales et al 2005;Gillardon et al 2007;Dani and Dencher 2008), particularly BN/SDS-PAGE to approach the protein 'interactome' in a near-native scenario (Krause 2006;Marques et al 2007). The mild solubilisation of proteins using non-ionic detergents, as involved in BN-PAGE helps to preserve the delicate assemblies of proteins like respiratory chain supercomplexes and ATP synthase oligomers.…”

Section: Introductionmentioning

confidence: 99%

Modulation of oxidative phosphorylation machinery signifies a prime mode of anti-ageing mechanism of calorie restriction in male rat liver mitochondria

et al. 2009

Self Cite

View full text Add to dashboard Cite

Mitochondria being the major source and target of reactive oxygen species (ROS) play a crucial role during ageing. We analyzed ageing and calorie restriction (CR)-induced changes in abundance of rat liver mitochondrial proteins to understand key aspects behind the age-retarding mechanism of CR. The combination of blue-native (BN) gel system with fluorescence Difference Gel Electrophoresis (DIGE) facilitated an efficient analysis of soluble and membrane proteins, existing as monomers or multi-protein assemblies. Changes in abundance of specific key subunits of respiratory chain complexes I, IV and V, critical for activity and/or assembly of the complexes were identified. CR lowered complex I assembly and complex IV activity, which is discussed as a molecular mechanism to minimize ROS production at mitochondria. Notably, the antioxidant system was found to be least affected. The GSH:GSSG couple could be depicted as a rapid mean to handle the fluctuations in ROS levels led by reversible metabolic shifts. We evaluated the relative significance of ROS generation against quenching. We also observed parallel and unidirectional changes as effect of ageing and CR, in subunits of ATP synthase, cytochrome P450 and glutathione S-transferase. This is the first report on such 'putatively hormetic' ageing-analogous effects of CR, besides the age-retarding ones.

show abstract

“…Comparative transcriptomic and proteomic studies have been initiated to determine global changes in mitochondria from young versus aged skeletal muscle [26,39,41,62,[65][66][67][68]. Native-difference gel electrophoresis (DIGE) is an approach that facilitates sensitive quantitative assessment of changes in membrane and soluble proteins.…”

Section: Proteomic Analysis Pertaining To Ageing Skeletal Musclementioning

confidence: 99%

Studies of Complex Biological Systems with Applications to Molecular Medicine: The Need to Integrate Transcriptomic and Proteomic Approaches

Silvestri¹,

Lombardi²,

Lange³

et al. 2014

Omics in Clinical Practice

View full text Add to dashboard Cite

Omics approaches to the study of complex biological systems with potential applications to molecular medicine are attracting great interest in clinical as well as in basic biological research. Genomics, transcriptomics and proteomics are characterized by the lack of an a priori definition of scope, and this gives sufficient leeway for investigators (a) to discern all at once a globally altered pattern of gene/protein expression and (b) to examine the complex interactions that regulate entire biological processes. Two popular platforms in "omics" are DNA microarrays, which measure messenger RNA transcript levels, and proteomic analyses, which identify and quantify proteins. Because of their intrinsic strengths and weaknesses, no single approach can fully unravel the complexities of fundamental biological events. However, an appropriate combination of different tools could lead to integrative analyses that would furnish new insights not accessible through one-dimensional datasets. In this review, we will outline some of the challenges associated with integrative analyses relating to the changes in metabolic pathways that occur in complex pathophysiological conditions (viz. ageing and altered thyroid state) in relevant metabolically active tissues. In addition, we discuss several new applications of proteomic analysis to the investigation of mitochondrial activity.

show abstract

Native‐DIGE: A new look at the mitochondrial membrane proteome

Cited by 19 publications

References 25 publications

Native electrophoretic techniques to identify protein–protein interactions

Native electrophoretic techniques to identify protein–protein interactions

Modulation of oxidative phosphorylation machinery signifies a prime mode of anti-ageing mechanism of calorie restriction in male rat liver mitochondria

Studies of Complex Biological Systems with Applications to Molecular Medicine: The Need to Integrate Transcriptomic and Proteomic Approaches

Contact Info

Product

Resources

About