shotgun-proteomics-sample-processing-automated-an-open-source-lab

Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot

Mass spectrometry-based shotgun proteomics experiments require multiple sample preparation steps, including enzymatic protein digestion and clean-up, ...

University of Colorado Anschutz Medical Campus

The heart is sensitive to oxidative damage but a global view on how the cardiac proteome responds to oxidative stressors has yet to fully emerge. Using quantitative tandem mass spectrometry, we assessed the effects of acute exposure of the oxidative stress inducer paraquat on protein expression in mouse hearts. We observed widespread protein expression changes in the paraquat-exposed heart especially in organelle-containing subcellular fractions. During cardiac response to acute oxidative stress, proteome changes are consistent with a rapid reduction of mitochondrial metabolism, coupled with activation of multiple antioxidant proteins, reduction of protein synthesis and remediation of proteostasis. In addition to differential expression, we saw evidence of spatial reorganizations of the cardiac proteome including the translocation of hexokinase 2 to more soluble fractions. Treatment with the antioxidants Tempol and MitoTEMPO reversed many proteomic signatures of paraquat but this reversal was incomplete. We also identified a number of proteins with unknown function in the heart to be triggered by paraquat, suggesting they may have functions in oxidative stress response. Surprisingly, protein expression changes in the heart correlate poorly with those in the lung, consistent with differential sensitivity or stress response in these two organs. The results and data set here could provide insights into oxidative stress responses in the heart and avail the search for new therapeutic targets.

Cody T. Thomas

Department of Medicine-Cardiology,
Consortium for Fibrosis Research & Translation,
School of Medicine,
Consortium for Fibrosis Research & Translation, School of Medicine

University of Colorado Anschutz Medical Campus

Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot

Cody T. Thomas

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medicine-Cardiology,Consortium for Fibrosis Research & Translation,School of Medicine,Consortium for Fibrosis Research & Translation, School of Medicine

University of Colorado Anschutz Medical Campus

Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot

Department of Medicine-Cardiology,
Consortium for Fibrosis Research & Translation,
School of Medicine,
Consortium for Fibrosis Research & Translation, School of Medicine