The Phosphocholine-binding Pocket on C-reactive Protein Is Necessary for Initial Protection of Mice against Pneumococcal Infection

Abstract

Background: CRP exerts antipneumococcal function in mice if administered during the early stages of pneumococcal infection. Results: CRP loses such antipneumococcal function when its phosphocholine-binding pocket is blocked. Conclusion:The phosphocholine-binding pocket on CRP participates in antipneumococcal function of CRP during the early stages of infection. Significance: Remodeling of CRP may be required for successful treatment of mice during the late stages of infection.

Purification of recombinant C-reactive protein mutants

“…PEt-affinity chromatography is used to purify those CRP mutants that have lost their PChbinding activity but not their PEt-binding activity (Agrawal et al, 2002;Gang et al, 2012). For example, CRP mutants F66A/E81A and F66A/T76Y/E81A do not bind PCh, but retain their PEt-binding activity.…”

“…PEt-conjugated Sepharose beads are prepared as follows (Gang et al, 2012). Twenty five ml of packed ECH-Sepharose 4B beads (GE Healthcare) are first washed with deionized water (pH 4.5) and then with 0.5 M NaCl.…”

“…The significance of the recognition function of CRP in its alternate structural pentameric conformation in host defense is unknown. However, ex vivo and in vivo experiments in animal models of inflammatory diseases indicate that CRP exerts, although limited, anti-pneumococcal, anti-atherosclerotic, anti-arthritic and antiamyloidogenic functions Agrawal et al, 2008Agrawal et al, , 2010Chang et al, 2012;Gang et al, 2012Gang et al, , 2015Jiang et al, 2006;Jones et al, 2011;Kovacs et al, 2007;Nayeri et al, 2010;Ngwa et al, 2016;Ozawa et al, 2016;Simons et al, 2014;Singh et al, 2008;Suresh et al, 2006Suresh et al, , 2007Szalai et al, 1995;Yother et al, 1982). CRP has also been shown to alleviate experimental autoimmune encephalomyelitis (Zhang et al, 2015).…”

“…For 2002; Gang et al, 2012Gang et al, , 2015Suresh et al, 2006Suresh et al, , 2007. Recombinant CRP mutants can be expressed in mammalian cells and, if expressed, can be purified from the cell culture media.…”

High Sensitivity C-Reactive Protein as Atherogenic Marker Among Type 2 Diabetes

“…C-reactive protein (CRP) is a 120 kDa acutephase protein which is produced mainly in the liver in response to IL-6 secretion (Ahmed, Jadhav, Hassan, & Meng, 2012), (Ferri et al, 2007). CRP is a member of the pentraxin family of proteins that binds to the Cpolysaccharide of Streptococcus pneumoniae (Gang et al, 2012).…”