Detection of Inositol Phosphates by Split PH Domains

Methods Mol Biol. 2020:2091:47-57. doi: 10.1007/978-1-0716-0167-9_4.

Abstract

The pleckstrin homology (PH) domain is a family of structurally conserved proteins which can bind inositol phosphate derivatives. Some proteins involved in cellular signaling and cytoskeletal organization possess split PH domains that assemble into a structure which can bind specific inositol phosphates. Here we describe the design of split PH domain from a structurally well-characterized PH domain of phospholipase C (PLC) δ1 and Bruton's tyrosine kinase (Btk), which selectively bind Ins(1,4,5)P3 and Ins(1,3,4,5)P4, respectively. The PH domains fold into a functional structure when the split halves are brought to close proximity, and can be utilized to detect specific inositol phosphate of interest.

Keywords: Fluorescence; Inositol phosphates; PH domain; Structure-based design.

MeSH terms

  • Agammaglobulinaemia Tyrosine Kinase / chemistry*
  • Hydrogen-Ion Concentration
  • Inositol 1,4,5-Trisphosphate / analysis
  • Inositol Phosphates / analysis*
  • Models, Molecular
  • Phospholipase C delta / chemistry*
  • Pleckstrin Homology Domains
  • Protein Conformation
  • Protein Folding

Substances

  • Inositol Phosphates
  • inositol-1,3,4,5-tetrakisphosphate
  • Inositol 1,4,5-Trisphosphate
  • Agammaglobulinaemia Tyrosine Kinase
  • Phospholipase C delta