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Crystallization and preliminary X‐ray crystallographic analysis of an artificial molten‐globular‐like triosephosphate isomerase protein of mixed phylogenetic origin
Author(s) -
Goyal Venuka Durani,
Yadav Pooja,
Kumar Ashwani,
Ghosh Biplab,
Makde Ravindra D.
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14020755
Subject(s) - triosephosphate isomerase , crystallization , crystallography , globular protein , phylogenetic tree , chemistry , globular cluster , biochemistry , enzyme , physics , organic chemistry , gene , quantum mechanics , galaxy
A bioinformatics‐based protein‐engineering approach called consensus design led to the construction of a chimeric triosephosphate isomerase (TIM) protein called ccTIM (curated consensus TIM) which is as active as Saccharomyces cerevisiae TIM despite sharing only 58% sequence identity with it. The amino‐acid sequence of this novel protein is as identical to native sequences from eukaryotes as to those from prokaryotes and shares some biophysical traits with a molten globular protein. Solving its crystal structure would help in understanding the physical implications of its bioinformatics‐based sequence. In this report, the ccTIM protein was successfully crystallized using the microbatch‐under‐oil method and a full X‐ray diffraction data set was collected to 2.2 Å resolution using a synchrotron‐radiation source. The crystals belonged to space group C 222 1 , with unit‐cell parameters a = 107.97, b = 187.21, c = 288.22 Å. Matthews coefficient calculations indicated the presence of six dimers in the asymmetric unit, with an approximate solvent content of 46.2%.