Deimination of arginine residues in nucleophosmin/B23 and histones in HL-60 granulocytes

Biochem Biophys Res Commun. 2002 Jan 25;290(3):979-83. doi: 10.1006/bbrc.2001.6303.

Abstract

Peptidylarginine deiminases (PADs) convert arginine residues in proteins into citrulline residues Ca(2+)-dependently. PAD V was recently found in granulocyte-differentiated HL-60 cells. To find a target of PAD V, we incubated HL-60 granulocytes with the calcium ionophore A23187 and studied deiminated proteins by immunocytochemistry and immunoblotting using a monospecific antibody to modified citrulline residues. Immunocytochemical signals were found in the nucleus upon incubation with A23187. Immunoblotting indicated that 40-, 18-, 17-, and 14-kDa proteins were preferentially deiminated. The 40-kDa protein, which was focused to pI 5.0 on two-dimensional gel electrophoresis, was identified as nucleophosmin/B23 by mass spectrometry. The 18-, 17-, and 14-kDa proteins extracted with 0.4 N H(2)SO(4) comigrated with histones H3, H2A, and H4, respectively, on two-dimensional gel electrophoresis specialized for histones. The citrulline content of histones amounted to about 10% of the histone molecules. We discuss the implications of deimination of these proteins for their nuclear functions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arginine / metabolism*
  • Calcimycin / pharmacology
  • Citrulline / analysis
  • Citrulline / immunology
  • Electrophoresis, Gel, Two-Dimensional
  • Granulocytes / metabolism*
  • Granulocytes / ultrastructure
  • HL-60 Cells
  • Histones / chemistry*
  • Histones / metabolism*
  • Humans
  • Hydrolases / metabolism
  • Immunoblotting
  • Immunohistochemistry
  • Ionophores / pharmacology
  • Kinetics
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / metabolism*
  • Protein Processing, Post-Translational
  • Protein-Arginine Deiminase Type 4
  • Protein-Arginine Deiminases

Substances

  • Histones
  • Ionophores
  • Nuclear Proteins
  • nucleophosmin
  • Citrulline
  • Calcimycin
  • Arginine
  • Hydrolases
  • PADI4 protein, human
  • Protein-Arginine Deiminase Type 4
  • Protein-Arginine Deiminases