Visualization of acetylated mitochondrial proteins in Swiss 3T3 cells. Acetylated mitochondrial-localized proteins were labeled with acetyl-lysine antibody (Cat. # AAC02) and alexa-488 secondary (green), or a fluorescent mitochondrial marker (mitotracker orange) shown in red.

Visualization of acetylated mitochondrial proteins in Swiss 3T3 cells. Acetylated mitochondrial-localized proteins were labeled with acetyl-lysine antibody (Cat. # AAC02) and alexa-488 secondary (green), or a fluorescent mitochondrial marker (mitotracker orange) shown in red.

Post-translational modification (PTM) crosstalk is recognized as a major cell-regulatory mechanism; a theory validated by in-depth, functional analysis of several well-investigated proteins. Recently, Horita et al. examined the PTM crosstalk that occurs in acetylated mitochondrial proteins in response to a mitochondrial stress-inducing agent, hydrogen peroxide (H2O2). Visual changes in the acetylated state of mitochondrial proteins were investigated by immunofluorescence and showed dynamic changes in response to H2O2. These investigators validated the acetylation state of 10 mitochondrial targets and also measured endogenous changes to the acetylation state of these proteins using Signal-Seeker acetyl-lysine detection tools. The endogenous acetylation (Ac), ubiquitination (Ub), SUMOylation 2/3 (SUMO 2/3), and tyrosine phosphorylation (pY) state of four target mitochondrial proteins were investigated with these toolkits to examine if PTM crosstalk commonly occurs on mitochondrial proteins. Each of the four proteins had unique PTM profiles, but diverging acetylation and ubiquitin or SUMO 2/3 signals appeared to be a common theme. Cytoskeleton’s pY, Ub, SUMO 2/3, and Ac Signal-Seeker PTM detection kits (Cat. # BK160, BK161, BK162, and BK163 respectively) were essential tools used in this study to examine endogenous and dynamic PTM crosstalk.

Link to citation:

Products used in this citation:

Signal-Seeker™ Phosphotyrosine Detection Kit (Cat. # BK160)

Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)

Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)

Signal-Seeker™ Acetyl-Lysine Detection Kit (Cat. # BK163)

BlastR Rapid Lysate Prep Kit (Cat. # BLR01)

Posted in News By

alex castellanos