Analysis of protein posttranslational modifications using DIGE-based proteomics.
Difference gel electrophoresis (DIGE) is most often used to assess relative changes in the expression levels of individual proteins in multiple complex samples, and this information is valuable in making inferences about relative protein activity. However, a protein's activity is not solely dependent upon its expression level. A change in activity may also be influenced by myriad posttranslational modifications (PTMs), including palmitoylation, ubiquitination, oxidation, and phosphorylation. In this chapter, we describe the use of DIGE to determine specific PTMs by introducing specific labels or changes in pI and/or molecular weight.
Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC, USA.
Robert M
RM
DeKroon
Jennifer B
JB
Robinette
Cristina
C
Osorio
Joseph S Y
JS
Jeong
Eric
E
Hamlett
Mihaela
M
Mocanu
Oscar
O
Alzate
D053000:Analytic Sample Preparation Methods; D007525:Isoelectric Focusing; D054878:Lipoylation; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling; D013379:Substrate Specificity; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*; D054875:Ubiquitination
D011506:Proteins