A proteomic analysis of C-reactive protein stimulated THP-1 monocytes

Abstract

BackgroundC-reactive protein (CRP) is a predictor of cardiovascular risk. It circulates as a pentameric protein in plasma. Recently, a potential dissociation mechanism from the disc-shaped pentameric CRP (pCRP) into single monomers (monomeric or mCRP) has been described. It has been shown that mCRP has strong pro-inflammatory effects on monocytes. To further define the role of mCRP in determining monocyte phenotype, the effects of CRP isoforms on THP-1 protein expression profiles were determined. The hypothesis to be tested was that mCRP induces specific changes in the protein expression profile of THP-1 cells that differ from that of pCRP.MethodsProtein cell lysates from control and mCRP, pCRP or LPS-treated THP-1 cells were displayed using 2-dimensional SDS PAGE and compared. Differentially expressed proteins were identified by MALDI-TOF MS and confirmed by Western blotting.ResultsmCRP significantly up-regulates ubiquitin-activating enzyme E1, a member of the ubiquitin-proteasome system in THP-1 monocytes. Furthermore, HSP 70, alpha-actinin-4 (ACTN4) and alpha-enolase/enolase 1 were upregulated. The proteomic profile of LPS and pCRP treated monocytes differ significantly from that of mCRP.ConclusionThe data obtained in this study support the hypothesis that isoform-specific effects of CRP may differentially regulate the phenotype of monocytes.

Characterization of globulin storage proteins of a low prolamin cereal species in relation to celiac disease

“…Total protein of 50 mg-s of crushed B. distachyon inbred line Bd21 seeds were extracted following the protocol of Dupont and co-workers13. Briefly, an SDS-Tris extraction buffer (pH 6.8) containing 2% SDS, 10% glycerol, 50 mM DTT and 40 mM Tris-HCl, pH 6.8 was used to extract the total protein at room temperature for one hour with a regular gentle vortexing.…”

Small heat shock proteins: recent developments

“…Mutations in these and other human sHSPs are associated with a wide range of diseases, the majority of which are neuropathies and myopathies (81)(82)(83)(84). Besides possible interactions with elements of the cytoskeleton (85)(86)(87), different members of the chaperone family are found to be present at sites of inflammation (88,89) and cellular defects like ischemic acute renal failure (90) or excitotoxic lesions (91). As a general protective effect, it was reported that high levels of sHSPs extend the life span of some organisms (92)(93)(94).…”

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

“…Therefore, scientists usually crosslink Igs captured on protein A or G via chemical crosslinkers such as dimethyl pimelimidate (DMP) and bis(sulfosuccinimidyl) suberate (BS3). [162–165] However, crosslinkers like DMP may also modify the amine groups on the antigen biding sites, leading to a decrease in antigen binding efficiency. [166] Konrad et.…”