Protein acyltransferase function of purified calreticulin. Part 1: characterization of propionylation of protein utilizing propoxycoumarin as the propionyl group donor

J Biochem. 2010 May;147(5):625-32. doi: 10.1093/jb/mvq002. Epub 2010 Jan 12.

Abstract

We have earlier reported that an endoplasmic reticulum luminal protein calreticulin (CR) mediated the acetylation of certain receptor proteins such as glutathione S-transferase (GST) by polyphenolic acetates, leading to irreversible inhibition. This function of calreticulin was termed calreticulin transacetylase. In this communication, we have demonstrated for the first time the ability of the purified recombinant calreticulin of a parasitic nematode Haemonchus contortus to transfer propionyl group from 7,8-Dipropoxy-4-methylcoumarin (DPMC) to recombinant Schistosoma japonicum glutathione S-transferase (rGST). Calreticulin transacetylase exhibited hyperbolic kinetics and yielded K(m) (140 microM) and V(max) (105 units) when the concentration of DPMC was varied keeping the concentration of rGST constant. rGST thus propionylated was found to positively interact with anti-acetyl lysine antibody. Also, the nanoscale LC-MS/MS analysis identified the propionylation sites on three lysine residues: Lys-11, -180 and -181 of rGST. These results highlight the transacylase function of calreticulin (CRTAase).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / metabolism*
  • Animals
  • Calreticulin / isolation & purification*
  • Calreticulin / metabolism*
  • Coumarins / chemistry*
  • Coumarins / metabolism*
  • Haemonchus / metabolism
  • Kinetics
  • Propionates / chemistry
  • Propionates / metabolism*
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • 7,8-dipropoxy-4-methylcoumarin
  • Calreticulin
  • Coumarins
  • Propionates
  • Recombinant Proteins
  • Acetyltransferases
  • propionic acid