Supported Cores

The Analytical Redox Core has now gained approval for an MUSC based formalized billing structure. The rate analysis was completed, formal user billing procedures are now implemented. The Core was also successful in the supplemental equipment grant application facilitating purchase of an automated high-content and high-throughput Biolog’s Phenotype MicroArray™ Technology (BIOLOG, Inc. Hayward, CA, USA). This technology leverages proprietary redox sensitive dyes to interrogate upwards of 1,400 pathways in living cells, in real time and will provide comprehensive “phenomics”. This platform will allow the interrogation of mitochondria function, substrate metabolism, sensitivity to drugs and toxins and mutations that impact bioenergetics. The ARB Core performed pilot data using the BiOLOG system and showed reductive stress alters mitochondrial structure and function, coordinate with changes in substrate utility in living cells. These data were presented to the External Advisory Board with enthusiasm to bring the technology to our research community. Previously, acquisition of the Phenotypic Microarray System has not been previously available to COBRE investigators. Core personnel continue with existing collaborations with Northwestern University. As in previous years, to accommodate this initiative, the COBRE EAB recommended increasing financial support of the ARB Core as appropriate. This was achieved by moving some of the funds from other sources. For Proteomics, Dr. Ball has now integrated her NIH shared instrumentation grant mass spectrometry capabilities.

 

The Proteomics Core has established methodology facilitating the characterization of redox-and drug-sensitive post-translational modifications of amino acids that are subject to modifications governed by changes in redox homeostasis. The core has been part of the COBRE since its inception and has proactively responded to the needs of investigators in focusing upon: cysteine modifications (e.g. sulfenic, sulfinic, sulfonic acid, S-glutathionylation, disulfide bonds); arginine (glycation by methylglyoxal, dihydroxyimidazolidine); tyrosine (nitrated and crosslinked), serine and threonine (phosphorylation); proline (oxidation); and lysine (acetylation, ubiquitin, glycation). Quantitative proteomic workflows utilizing metabolic labeling (SILAC), multiplexed isobaric tagging (TMT), and label free quantitation (MQLFQ) for differential protein expression, identification of differentially regulated sites of modification, and protein interactions are routinely performed. The objective of the Core is to continue to update and improve LC-MS/MS analyses for discovery and targeted proteomics and has been upgraded for this Phase III application by including state-of-the-art equipment and technologies to provide access to MALDI-Imaging machines for biomarker discovery. Dr. Lauren Ball is an Associate Professor in the Dept. Of Pharmacology and has served as Core Director of the Redox Center Proteomics Core and the Mass Spectrometry Facility shared resource at MUSC since the COBRE beginning. Dr. Ball has considerable expertise in analytical proteomics and partly as a direct consequence of her work on the COBRE, has specifically established expertise and a national reputation in redox based proteomic techniques.

 

The Cell & Molecular Redox Imaging has enabled the CMRI to provide the infrastructure, expertise and training to support the application of the latest imaging techniques for the visualization of subject matter relevant to novel redox biology and pathobiology. Lemasters is an internationally recognized expert in the role of mitochondria in pathobiology, especially in relation to oxidative stress, ethanol/drug-induced hepatic injury, ischemia-reperfusion (IR) injury, organ preservation for transplantation, and mitochondrial autophagy (mitophagy). He was recruited to MUSC from the University of North Carolina in 2006 to fulfill the need of MUSC investigators to have access to state-of-the-art capabilities in biological imaging and has been part of the COBRE since its inception. The facility emphasizes advanced and innovative imaging capabilities to COBRE investigators, as well as hands-on and didactic training in advanced imaging. As outlined in the Core description, a variety of new equipment has recently been purchased (some with funds from the administrative supplement) and incorporated, allowing greater flexibility in the services offered.