Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans

J Microbiol. 2016 Jun;54(6):440-4. doi: 10.1007/s12275-016-6089-5. Epub 2016 May 27.

Abstract

Chaperonins (CPNs) are megadalton sized ATP-dependent nanomachines that facilitate protein folding through complex cycles of complex allosteric articulation. They consist of two back-to-back stacked multisubunit rings. CPNs are usually classified into Group I and Group II. Here, we report the crystallization of both the AMPPNP (an ATP analogue) and ADP bound forms of a novel CPN, classified as belonging to a third Group, recently discovered in the extreme thermophile Carboxydothermus hydrogenoformans. Crystals of the two forms were grown by the vapor batch crystallization method at 295 K. Crystals of the Ch-CPN/AMPPNP complex diffracted to 3.0 Å resolution and belonged to the space group P422, with unit-cell parameters a = b = 186.166, c = 160.742 Å. Assuming the presence of four molecules in the asymmetric unit, the solvent content was estimated to be about 60.02%. Crystals of the Ch-CPN/ADP complex diffracted to 4.0 Å resolution and belonged to the space group P4212, with unit-cell parameters a = b = 209.780, c = 169.813Å. Assuming the presence of four molecules in the asymmetric unit, the solvent content was estimated to be about 70.19%.

Keywords: AT-Pfueled nanomachine; Group III chaperonins; crystallization; protein folding.

MeSH terms

  • Adenosine Diphosphate / chemistry
  • Adenosine Diphosphate / metabolism
  • Adenosine Triphosphate / chemistry
  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Bacterial Proteins / metabolism
  • Chaperonins / chemistry*
  • Chaperonins / genetics
  • Chaperonins / isolation & purification*
  • Chaperonins / metabolism
  • Crystallography, X-Ray
  • Models, Molecular
  • Molecular Sequence Data
  • Sequence Alignment
  • Thermoanaerobacterium / chemistry
  • Thermoanaerobacterium / genetics
  • Thermoanaerobacterium / metabolism*

Substances

  • Bacterial Proteins
  • Adenosine Diphosphate
  • Adenosine Triphosphate
  • Chaperonins