Development of phosphorylation site-specific antibodies to nuclear receptors

Methods Mol Biol. 2009:505:221-35. doi: 10.1007/978-1-60327-575-0_13.

Abstract

Protein phosphorylation is a versatile posttranslational modification that can regulate nuclear receptor function. Although the precise role of receptor phosphorylation is not fully understood, it appears that it functions to direct or refine receptor activity in response to particular physiological requirements. Identifying and characterizing specific nuclear receptor phosphorylation sites is an important step in elucidating the role(s) receptor phosphorylation plays in function. Although traditional methods of metabolic labeling and in vitro protein phosphorylation have been informative, receptor phosphorylation site-specific antibodies are simple and reliable tools to study receptor phosphorylation. This chapter will discuss how to develop nuclear receptor phosphorylation site-specific antibodies to elucidate function.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Phospho-Specific* / chemistry
  • Antibodies, Phospho-Specific* / metabolism
  • Antibody Specificity
  • Biological Assay / methods
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Humans
  • Liver X Receptors
  • Molecular Sequence Data
  • Orphan Nuclear Receptors
  • Phosphorylation
  • Protein Engineering / methods*
  • Receptors, Cytoplasmic and Nuclear / chemistry*
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Reproducibility of Results

Substances

  • Antibodies, Phospho-Specific
  • DNA-Binding Proteins
  • Liver X Receptors
  • Orphan Nuclear Receptors
  • Receptors, Cytoplasmic and Nuclear