Analytical Redox Biology Core

Core Director: Danyelle Townsend, Ph.D.

Understanding the complexities of redox mediated signaling events requires a multidisciplinary approach.  The SC COBRE in Oxidants, Redox Balance and Stress Signalling has assembled a cohort of promising junior faculty with expertise in relevant biomedical model systems. Analytical biochemistry specific to the detection and quantification of redox sensitive molecules and coordinate protein changes that drive homeostasis is a unique niche fulfilled by the Analytical Redox Biology Core (ARBC).

The primary objective of the Core is to provide comprehensive analytical redox biochemistry methods and mentoring support for the COBRE junior faculty with the goal to advance their research endeavors, publications and fundability.  The specific aims of the ARBC are: 1) Provide ROS /RNS identification and quantification using state-of-the-art techniques;  2) Perform quantitative analysis of ROS/RNS (redox molecules and metabolites), including those associated with calcium mobilization and changes in energy metabolism; 3) Provide expertise and technology for in depth biochemical analysis of thiol-centered enzyme activities and define protein:protein interactions.

Since oxidative (nitrosative) stress often is associated with a conditional increase in antioxidant protection, the Core has established methods to detect and measure various antioxidant enzyme activities as a function of oxidant stress/antioxidant protection equilibrium. Comprehensive analysis of redox status also includes measurement of intracellular GSH, GSSG, protein surface and “buried” thiols utilizing both endpoint and/ or real-time kinetic measurements with millisecond resolution. In complex studies of redox signaling, certain protein:protein interactions appear to be redox dependent and attributed to post-translational modifications, including S-nitrosylation and S-glutathionylation. The ARBC has developed fluorescent labeling and FRET analysis to evaluate redox dependent protein:protein interactions with subsequent in silico molecular modeling using ZDOCK, GOLD Suite (v 5.2) software. Collectively, these technologies will provide a multidisciplinary approach to advance the understanding of redox mediated signaling events specific to the model systems presented by the junior faculty in their research.

Analytical biochemistry specific to the detection and quantification of redox sensitive molecules and coordinate protein changes that drive homeostasis is a unique niche that was earlier provided by a dedicated Bioenergetics Core. The untimely death of Dr. Craig Beeson led to the decision that such technologies should be assimilated into the ARC and remain available to the members of the SC COBRE in Oxidants, Redox Balance and Stress Signaling and the greater scientific community. The ARC now supports this scaled-down focus on and bioenergetics for redox biology research by leveraging the existing resources of the MUSC Bioenergetics Core, the day to day running of which is maintained by Mrs. Beeson. The MUSC Bioenergetics Core is a multi-user shared instrumentation resource actively utilized by faculty level investigators and their trainees throughout MUSC. In such leveraging, the ARC benefits not only COBRE investigators but the MUSC research community overall by providing expert advice, consultation, and education in bioenergetic profiling and quantitative assessment of redox homeostasis. We now include these services within ARC using a user-fee approach to Seahorse usage.