Purification, crystallization and preliminary crystallographic analysis of a ribosome-recycling factor fromThermoanaerobacter tengcongensis(TteRRF)

Abstract

Ribosome-recycling factor (RRF) plays an essential role in the fourth step of protein synthesis in prokaryotes. RRF combined with elongation factor G (EF-G) disassembles the post-termination ribosome complex and recycles the protein synthesis machine for the next round of translation. A reductive-methylation-modified RRF fromThermoanaerobacter tengcongensis(TteRRF) has been crystallized using the vapour-diffusion method. The crystal grew in a condition consisting of 0.1 Mcitric acid pH 3.5, 3.0 MNaCl and 50 mg ml−1methylated protein solution at 289 K. A complete data set was collected from a crystal to 2.80 Å resolution using synchrotron radiation at 100 K. The crystal belonged to space groupP6122/P6522 with unit-cell parametersa=b= 103.26,c= 89.17 Å. The asymmetric unit was estimated to contain one molecule ofTteRRF.