Crystallization and Preliminary X-ray Characterization of the Genetically Encoded Fluorescent Calcium Indicator Protein GCaMP2

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Jul 1;64(Pt 7):629-31. doi: 10.1107/S1744309108016059. Epub 2008 Jun 11.

Abstract

Fluorescent proteins and their engineered variants have played an important role in the study of biology. The genetically encoded calcium-indicator protein GCaMP2 comprises a circularly permuted fluorescent protein coupled to the calcium-binding protein calmodulin and a calmodulin target peptide, M13, derived from the intracellular calmodulin target myosin light-chain kinase and has been used to image calcium transients in vivo. To aid rational efforts to engineer improved variants of GCaMP2, this protein was crystallized in the calcium-saturated form. X-ray diffraction data were collected to 2.0 A resolution. The crystals belong to space group C2, with unit-cell parameters a = 126.1, b = 47.1, c = 68.8 A, beta = 100.5 degrees and one GCaMP2 molecule in the asymmetric unit. The structure was phased by molecular replacement and refinement is currently under way.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / chemistry*
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / genetics*
  • Crystallization
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / genetics*
  • Indicators and Reagents
  • Molecular Sequence Data
  • X-Ray Diffraction*

Substances

  • Indicators and Reagents
  • Green Fluorescent Proteins
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2