Scientists from Vanderbilt University and the Universidad de la República in Uruguay, in collaboration with other research centers, have successfully resurrected a "dead antibody" to study the mysteries of cytochrome c. This versatile protein is an essential component of the cell's energy-generating capacity and plays a major role in programmed cell death or apoptosis. The research team's achievement opens new avenues for understanding cytochrome c's alternate conformations, which have long evaded scientific scrutiny.
Background on Cytochrome c
Cytochrome c is a protein found primarily in the inner membrane of mitochondria, where it assists in energy production. However, when it's released to the inner cellular fluid or cytosol, it plays a crucial role in the regulated death of cells that are no longer required. Additionally, scientists have detected cytochrome c in secretory and bacterial vesicles, implying that it may be involved in undiscovered functions.
The Resurrection of a 'Dead Antibody'
In 2009, Uruguayan scientist Rafael Radi and his collaborators discovered a new, tridimensional structure of cytochrome c using a unique monoclonal antibody. However, the hybridoma that produced the antibody was lost when the liquid nitrogen tank storing it malfunctioned. Attempts to rebuild the hybridoma were unsuccessful. Ten years later, technological advancements enabled Radi's team, in collaboration with Vanderbilt colleagues and other research centers, to determine the antibody's full amino-acid sequence and recreate it in the laboratory.
Unique Tool for Protein Dynamics and Plasticity
The researchers published their findings in the Proceedings of the National Academy of Sciences, demonstrating that the resurrected antibody detected alternate conformations of cytochrome c in living cells. These conformations may play a role in regulating gene expression, and the antibody could become a unique tool for studying protein dynamics and plasticity.
Collaboration and Synergy of Great Science
Radi's work has defined the role of free radicals, oxidants, and mitochondrial dysfunction in human disease. He is a professor and chair of Biochemistry at the Facultad de Medicina, Universidad de la República, and an International Member of the U.S. National Academy of Sciences. He is also an adjunct member of the Vanderbilt University faculty in the Department of Biochemistry and Vanderbilt Institute for Chemical Biology. According to Associate Director of the Vanderbilt Vaccine Center James Crowe, "This work illustrates the synergy of great science and collaborative spirit that exemplifies Vanderbilt core labs."
Journal Information: Florencia Tomasina et al, De novo sequencing and construction of a unique antibody for the recognition of alternative conformations of cytochrome c in cells, Proceedings of the National Academy of Sciences (2022). DOI: 10.1073/pnas.2213432119