title: On-Membrane Renaturation of Recombinant Ro60 Autoantigen by Calcium Ions.
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.
authors.affiliation: Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA, biji-kurien@omrf.org.
authors.forename: Biji T
authors.initials: BT
authors.lastname: Kurien
authors.forename: Michael P
authors.initials: MP
authors.lastname: Bachmann
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D002118:Calcium / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D008567:Membranes, Artificial; D011145:Polyvinyls / Q000737:chemistry*; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000032:analysis; D012261:Ribonucleoproteins / Q000032:analysis*; D000099207:SS-A Antigen; D005465:Fluorocarbon Polymers
chemical_list: D002118:Calcium; D008567:Membranes, Artificial; D011145:Polyvinyls; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers