Phosphorylation of A170 stress protein by casein kinase II-like activity in macrophages

Biochem Biophys Res Commun. 1997 Dec 8;241(1):157-63. doi: 10.1006/bbrc.1997.7783.

Abstract

A170 is an oxidative stress-inducible protein having a Zinc finger domain, two PEST sequences, and many potential phosphorylation sites for serine/threonine kinases. These structural features suggest that the phosphorylation of A170 affects its function and degradation. We have found that A170 is phosphorylated in cultured murine peritoneal macrophages. In addition, using recombinant A170 proteins, we found two proteins of 40 and 44 kDa with kinase activity in cell extracts using an in-gel kinase assay. We compared the properties of the intrinsic A170 kinases with those of mitogen-activated protein kinase (ERK 2), protein kinase A (PKA), casein kinase II (CK II), and protein kinase C, since their catalytic subunits have molecular masses similar to A170 kinases. ERK 2, CK II, and PKA phosphorylated recombinant A170 as a substrate. The 40 and 44 kDa kinases present in the macrophage extract were similar to alpha and alpha' subunits of CK II in respect to substrate specificity, pharmacological properties, immuno-reactivities, and ubiquitous expression in tissues.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Animals
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Casein Kinase II
  • Caseins / metabolism
  • Cells, Cultured
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Female
  • Guanosine Triphosphate / pharmacology
  • Heat-Shock Proteins / metabolism*
  • Heparin / pharmacology
  • Macrophages, Peritoneal / enzymology*
  • Mice
  • Mice, Inbred Strains
  • Mitogen-Activated Protein Kinase 1
  • Molecular Weight
  • Myelin Basic Protein / metabolism
  • Phosphorylation
  • Polymerase Chain Reaction
  • Protein Serine-Threonine Kinases / chemistry
  • Protein Serine-Threonine Kinases / isolation & purification
  • Protein Serine-Threonine Kinases / metabolism*
  • Recombinant Proteins / metabolism
  • Sequestosome-1 Protein
  • Substrate Specificity

Substances

  • Adaptor Proteins, Signal Transducing
  • Caseins
  • Heat-Shock Proteins
  • Myelin Basic Protein
  • Recombinant Proteins
  • Sequestosome-1 Protein
  • Sqstm1 protein, mouse
  • Guanosine Triphosphate
  • Heparin
  • Casein Kinase II
  • Protein Serine-Threonine Kinases
  • Cyclic AMP-Dependent Protein Kinases
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinase 1