Isolation of nucleic acid binding proteins: an approach for isolation of cell surface, nucleic acid binding proteins

Ann N Y Acad Sci. 2004 Jun:1022:239-43. doi: 10.1196/annals.1318.037.

Abstract

An approach for isolation of cell surface, nucleic acid binding proteins is described. This approach relies on affinity modification of the proteins of living cells with reactive oligonucleotides bearing a haptenic group. Covalently modified proteins were isolated by hapten-specific affinity chromatography with subsequent SDS-PAGE. Isolated 68-kDa proteins responsible for the binding of oligonucleotides were MS/MS sequenced and identified as keratin K1, keratin K10, keratin K2e, and albumin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Squamous Cell / pathology
  • Cell Line, Tumor
  • Chromatography, Affinity
  • DNA-Binding Proteins / isolation & purification*
  • DNA-Binding Proteins / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Haptens / chemistry
  • Humans
  • Keratin-10
  • Keratin-2
  • Keratins / chemistry
  • Keratins / isolation & purification
  • Keratins / metabolism
  • Mass Spectrometry
  • Molecular Weight
  • Neoplasms, Squamous Cell / pathology
  • Oligodeoxyribonucleotides / chemical synthesis
  • Oligodeoxyribonucleotides / chemistry
  • Oligodeoxyribonucleotides / isolation & purification*
  • Oligodeoxyribonucleotides / metabolism*
  • Sequence Analysis, Protein
  • Serum Albumin / isolation & purification
  • Serum Albumin / metabolism

Substances

  • DNA-Binding Proteins
  • Haptens
  • KRT10 protein, human
  • KRT2 protein, human
  • Keratin-2
  • Oligodeoxyribonucleotides
  • Serum Albumin
  • Keratin-10
  • Keratins