Monitoring cytoplasmic protein complexes with blue native gel electrophoresis and stable isotope labelling with amino acids in cell culture: Analysis of changes in the 20S proteasome

2011 ◽  
Vol 32 (14) ◽  
pp. 1819-1823 ◽  
Author(s):  
Timothy A. Couttas ◽  
Mark J. Raftery ◽  
Melissa A. Erce ◽  
Marc R. Wilkins
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Gel Electrophoresis ◽  
Protein Complexes ◽  
20S Proteasome ◽  
Cytoplasmic Protein ◽  
Stable Isotope Labelling ◽  
Isotope Labelling ◽  
Native Gel ◽  
Blue Native

scholarly journals Dynamics of murine brain protein synthesis in vivo identify the hippocampus, cortex and cerebellum as highly active metabolic sites

2019 ◽  
Author(s):  
Ser Sue Ng ◽  
Jung Eun Park ◽  
Wei Meng ◽  
Christopher Li-Hsian Chen ◽  
Raj N. Kalaria ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Protein Synthesis ◽  
Stable Isotope ◽  
Density Lipoprotein ◽  
Apolipoprotein A1 ◽  
Stable Isotope Labelling ◽  
Isotope Labelling ◽  
Highly Active

AbstractIdentification of proteins that are synthesized de novo in response to specific microenvironmental cues is critical to understanding the molecular mechanisms that underpin key physiological processes and pathologies. Here we report that a brief period of pulsed SILAC diet (Stable Isotope Labelling by Amino acids in Cell culture) enables determination of biological functions corresponding to actively translating proteins in the mouse brain. Our data demonstrate that the hippocampus, cortex and cerebellum are highly active sites of protein synthesis, rapidly expressing key mediators of nutrient sensing and lipid metabolism, as well as critical regulators of synaptic function, axon guidance, and circadian entrainment. Together, these findings confirm that protein metabolic activity varies significantly between brain regions in vivo and indicate that pSILAC-based approaches can identify specific anatomical sites and biological pathways likely to be suitable for drug targeting in neurodegenerative disorders.AbbreviationsApoA1: Apolipoprotein A1, ApoA4: Apolipoprotein A4, ApoE: Apolipoprotein E, ApoJ/Clu: Apolipoprotein J/Clusterin, App: Amyloid-β precursor/A4 protein: App, HDL: high density lipoprotein, Lrp1: Low density lipoprotein receptor-related protein 1, pSILAC: pulsed SILAC, pSIVOM: pulsed-SILAC in vivo labelling in mouse, SILAC: Stable Isotope Labelling by Amino acids in Cell culture)

Blue-Native Gel Electrophoresis for the Characterization of Protein Complexes in Plants

2006 ◽  
pp. 343-352 ◽  
Author(s):  
Jesco Heinemeyer ◽  
Dagmar Lewejohann ◽  
Hans-Peter Braun
Keyword(s):  
Gel Electrophoresis ◽  
Protein Complexes ◽  
Native Gel ◽  
Blue Native
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