title: C-reactive protein (CRP) aptamer binds to monomeric but not pentameric form of CRP.
abstract: Native C-reactive protein (CRP) is composed of five identical subunits arranged in a pentameric structure (pCRP). Binding of pCRP to damaged cell membranes produces a second isoform, modified CRP, which has similar antigenicity to isolated monomeric subunits of CRP (mCRP). Emerging evidence indicates that modified CRP plays a role in inflammation and atherosclerosis, however, there are very few techniques that can distinguish the different isoforms of CRP. Here we show that an RNA aptamer binds specifically to mCRP and not to pCRP. Using this aptamer, we describe a simple, fast, and sensitive assay to detect nanomolar concentrations of mCRP using fluorescence anisotropy. In addition, we show that this aptamer can be used to detect mCRP in polyacrylamide gels and bound to a surface using total internal reflection fluorescence microscopy. The biological activity of the mCRP we prepared by heating pCRP with 0.1% sodium dodecyl sulfate was confirmed by observing binding to the complement protein, C1q. This probe provides an important tool for CRP research and has the potential to improve clinical diagnostics that predict risk for cardiovascular disease.
authors: Department of Chemistry, University of Colorado Denver, Denver, CO 80217-3364, USA.
authors: Min S
authors: MS
authors: Wang
authors: Joshua C
authors: JC
authors: Black
authors: Michelle K
authors: MK
authors: Knowles
authors: Scott M
authors: SM
authors: Reed
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism*; D002097:C-Reactive Protein / Q000737:chemistry* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005454:Fluorescence Polarization; D006801:Humans; D057230:Limit of Detection; D011485:Protein Binding; D020033:Protein Isoforms / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors
chemical_list: D052157:Aptamers, Nucleotide; D020033:Protein Isoforms; D002097:C-Reactive Protein