Abstract
Calcium plays an important role in many biochemical processes in different cell types. This divalent cation interacts with specific calcium-binding proteins that serve as calcium sensors and regulatory proteins to mediate its function. Previously, we found that calcium was involved in the protein-protein interaction observed between Ro60 multiple antigenic peptides and Ro60 autoantigen. Since calcium bound Ro60 multiple antigenic peptides, we hypothesized that it would renature human recombinant Ro60 on a protein blot. We found that anti-Ro60 antibodies bound significantly higher to the recombinant Ro60 antigen that was incubated with calcium compared to that incubated without calcium on a polyvinylidene fluoride (PVDF) blot. Since the immunological epitopes of Ro60 are mainly conformational, we believe that calcium induced a more native tertiary structure in recombinant Ro60 autoantigen following blotting to a PVDF membrane.
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Acknowledgement
We express our thanks to Natalie B. Reed and Alexandra Taylor for their excellent technical assistance.
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Kurien, B.T., Bachmann, M.P. (2015). On-Membrane Renaturation of Recombinant Ro60 Autoantigen by Calcium Ions. In: Kurien, B., Scofield, R. (eds) Detection of Blotted Proteins. Methods in Molecular Biology, vol 1314. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2718-0_26
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DOI: https://doi.org/10.1007/978-1-4939-2718-0_26
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2717-3
Online ISBN: 978-1-4939-2718-0
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