Identification of apoptosis-associated proteins in a human Burkitt lymphoma cell line. Cleavage of heterogeneous nuclear ribonucleoprotein A1 by caspase 3

J Biol Chem. 1998 Oct 23;273(43):28057-64. doi: 10.1074/jbc.273.43.28057.

Abstract

Apoptosis or programmed cell death is essential in the process of controlling lymphocyte growth and selection. We identified proteins that are involved in anti-IgM antibody-mediated apoptosis using a subclone of the human Burkitt lymphoma cell line BL60. Apoptosis-associated proteins were detected by high resolution two-dimensional gel electrophoresis on a micropreparative scale. Comparison of the high resolution two-dimensional gel electrophoresis protein patterns from apoptotic and non-apoptotic cells showed differences in approximately 80 spots including protein modifications. Analysis of the predominantly altered proteins was performed by internal Edman microsequencing and/or by peptide mass fingerprinting using matrix-assisted laser desorption/ionization mass spectrometry. Analysis was significantly improved by using new micropreparative high resolution two-dimensional gels employing high protein concentrations. The following 12 apoptosis-associated proteins were identified: heterogeneous nuclear ribonucleoprotein (hnRNP) A1, hnRNP C1/C2, FUSE-binding protein, dUTPase, lymphocyte-specific protein LSP1, UV excision repair protein RAD23 homologue B (HHR23B), 60 S acidic ribosomal protein P0 (L10E), heterochromatin protein 1 homologue alpha (HP1alpha), nucleolin, lamin, neutral calponin, and actin. Fragmentation of actin, hnRNP A1, hnRNP C1/C2, 60 S acidic ribosomal protein P0, lamin, and nucleolin could be inhibited by benzyloxycarbonyl-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone, a selective irreversible inhibitor of CPP32 (caspase 3).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Burkitt Lymphoma / metabolism*
  • Calcium-Binding Proteins / metabolism
  • Caspase 3
  • Caspase Inhibitors
  • Caspases / metabolism*
  • Chromobox Protein Homolog 5
  • Chromosomal Proteins, Non-Histone / metabolism
  • DNA Repair Enzymes
  • DNA-Binding Proteins / metabolism
  • Electrophoresis, Gel, Two-Dimensional
  • Heterogeneous Nuclear Ribonucleoprotein A1
  • Heterogeneous-Nuclear Ribonucleoprotein Group A-B*
  • Heterogeneous-Nuclear Ribonucleoprotein Group C*
  • Heterogeneous-Nuclear Ribonucleoproteins
  • Humans
  • Immunoglobulin M
  • Immunologic Capping
  • Immunomagnetic Separation
  • Lamins
  • Microfilament Proteins
  • Neoplasm Proteins / metabolism*
  • Nuclear Proteins / metabolism
  • Nucleolin
  • Oligopeptides / pharmacology
  • Phosphoproteins / metabolism
  • Pyrophosphatases / metabolism
  • RNA, Heterogeneous Nuclear
  • RNA-Binding Proteins / metabolism
  • Receptors, Antigen, B-Cell
  • Ribonucleoproteins / metabolism*
  • Ribosomal Proteins / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tumor Cells, Cultured

Substances

  • C1 HNRNP
  • CBX5 protein, human
  • Calcium-Binding Proteins
  • Caspase Inhibitors
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • HNRNPC protein, human
  • Heterogeneous Nuclear Ribonucleoprotein A1
  • Heterogeneous-Nuclear Ribonucleoprotein Group A-B
  • Heterogeneous-Nuclear Ribonucleoprotein Group C
  • Heterogeneous-Nuclear Ribonucleoproteins
  • Immunoglobulin M
  • LSP1 protein, human
  • Lamins
  • Microfilament Proteins
  • Neoplasm Proteins
  • Nuclear Proteins
  • Oligopeptides
  • Phosphoproteins
  • RAD23B protein, human
  • RNA, Heterogeneous Nuclear
  • RNA-Binding Proteins
  • Receptors, Antigen, B-Cell
  • Ribonucleoproteins
  • Ribosomal Proteins
  • benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone
  • hnRNPA1 protein, human
  • ribosomal protein P0
  • Chromobox Protein Homolog 5
  • RAD23A protein, human
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Pyrophosphatases
  • dUTP pyrophosphatase
  • DNA Repair Enzymes