Purification and determination of the modifying protein responsible for the post-synthetic modification of creatine kinase (EC 2.7.3.2) and enolase(EC 4.2.1.11)

Abstract

SummaryThe purification of a serum protein, responsible for the postsynthetic modification of CK and enolase, is described. A purification of about 1300-fold could be reached after subsequent chromatography of human serum on DEAE cellulose and Sephacryl S-200 Superfine followed by affinity chromatography using antibodies against human serum albumin, C3 and C4 and against total human serum proteins. A recovery of 160% of modifying activity was found. The molecular mass and the isoelectric point of the modifying protein have been determined.It is concluded that the concentration of the modifying protein in human serum is < 210 mg/l.

Biomarkers of Cardiac Injury: An Update

“…71,72 Enolase also undergoes postsynthetic modifications like CKMM. 73 At present, it appears that the measurement of β-enolases will not be able to differentiate between skeletal and myocardial muscle damage. An assay more specific for αβ-enolase might be more useful.…”

State and diagnostic value of plasma tissue factor in early‐hospitalised patients with chest pain

“…However, this possibility is excluded by the mentioned quantitative agreement between the TF antigen concentrations found in the present study and those reported in the studies quoted above for a variety of ELISAs. Apparently, the bulk of plasma TF is protected from the proteolytic fragmentation demonstrated for circulating tissue proteins, such as creatine kinase and enolase (Van Landeghem et al , 1985;), or TnT (Diris et al , 2004). Due to the lack of a membrane‐binding region, soluble TF has only a few percent of the procoagulant activity of lipidated full‐length TF (Waxman et al , 1992) and the same is true for the alternatively spliced human TF variant (ashTF) recently found by Bogdanov et al (2003).…”