Crystallization and preliminary X-ray crystallographic analysis of the PH-GRAM domain of human MTMR4

Acta Crystallogr F Struct Biol Commun. 2014 Sep;70(Pt 9):1280-3. doi: 10.1107/S2053230X14017658. Epub 2014 Aug 27.

Abstract

Phosphoinositide lipid molecules play critical roles in intracellular signalling pathways and are regulated by phospholipases, lipid kinases and phosphatases. In particular, phosphatidylinositol 3-phosphate and phosphatidylinositol 3,5-bisphosphate are related to endosomal trafficking events through the recruitment of effector proteins and are involved in the degradation step of autophagy. Myotubularin-related proteins (MTMRs) are a large family of phosphatases that catalyze the dephosphorylation of phosphatidylinositol 3-phosphate and phosphatidylinositol 3,5-bisphosphate at the D3 position, thereby regulating cellular phosphoinositide levels. In this study, the PH-GRAM domain of human MTMR4 was cloned, overexpressed in Escherichia coli, purified and crystallized by the vapour-diffusion method. The crystals diffracted to 3.20 Å resolution at a synchrotron beamline and belonged to either space group P61 or P65, with unit-cell parameters a = b = 109.10, c = 238.97 Å.

Keywords: MTMR4; PH-GRAM domain; myotubularin-related proteins; phosphatase; phosphoinositide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Crystallization
  • Crystallography, X-Ray / methods*
  • DNA Primers
  • Humans
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Tyrosine Phosphatases, Non-Receptor / chemistry*

Substances

  • DNA Primers
  • MTMR4 protein, human
  • Protein Tyrosine Phosphatases, Non-Receptor