title: Urea-induced denaturation of apolipoprotein serum amyloid A reveals marginal stability of hexamer.
abstract: Serum Amyloid A (SAA) is an acute phase reactant protein that is predominantly found bound to high-density lipoprotein in plasma. Upon inflammation, the plasma concentration of SAA can increase dramatically, occasionally leading to the development of amyloid A (AA) amyloidosis, which involves the deposition of SAA amyloid fibrils in major organs. We previously found that the murine isoform SAA2.2 exists in aqueous solution as a hexamer containing a central channel. Here we show using various biophysical and biochemical techniques that the SAA2.2 hexamer can be totally dissociated into monomer by approximately 2 M urea, with the concerted loss of its alpha-helical structure. However, limited trypsin proteolysis experiments in urea showed a conserved digestion profile, suggesting the preservation of major backbone topological features in the urea-denatured state of SAA2.2. The marginal stability of hexameric SAA2.2 and the presence of residual structure in the denatured monomeric protein suggest that both forms may interconvert in vivo to exert different functions to meet the various needs during normal physiological conditions and in response to inflammatory stimuli.
authors.affiliation: Rensselaer Polytechnic Institute, Department of Chemistry and Chemical Biology, Troy, NY 12180, USA.
authors.forename: Limin
authors.initials: L
authors.lastname: Wang
authors.forename: Wilfredo
authors.initials: W
authors.lastname: Colón
mesh_terms: D000818:Animals; D001704:Biopolymers / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D051379:Mice; D017433:Protein Structure, Secondary / Q000187:drug effects*; D000685:Serum Amyloid A Protein / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014357:Trypsin / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology*
chemical_list: D001704:Biopolymers; D003432:Cross-Linking Reagents; C489995:Saa2 protein, mouse; D000685:Serum Amyloid A Protein; D014508:Urea; D014357:Trypsin