Cloning, preliminary characterization and crystallization of nucleoside hydrolases from Caenorhabditis elegans and Campylobacter jejuni.

TitleCloning, preliminary characterization and crystallization of nucleoside hydrolases from Caenorhabditis elegans and Campylobacter jejuni.
Publication TypeJournal Article
Year of Publication2003
AuthorsVersées, W., E. Van Holsbeke, S. De Vos, K. Decanniere, I. Zegers, and J. Steyaert
JournalActa Crystallogr D Biol Crystallogr
Volume59
IssuePt 6
Pagination1087-9
Date Published2003 Jun
ISSN0907-4449
KeywordsAnimals, Caenorhabditis elegans, Campylobacter jejuni, Cloning, Molecular, Crystallization, Crystallography, X-Ray, Escherichia coli, Kinetics, N-Glycosyl Hydrolases, Reverse Transcriptase Polymerase Chain Reaction
Abstract

The nucleoside hydrolases (NHs) are a family of nucleoside-modifying enzymes. They play an important role in the purine-salvage pathway of many pathogenic organisms which are unable to synthesize purines de novo. Although well characterized in protozoan parasites, their precise function and mechanism remain unclear in other species. For the first time, NHs from Caenorhabditis elegans and Campylobacter jejuni, which are representatives of mesozoa and bacteria, respectively, have been cloned and purified. Steady-state kinetics indicate a different substrate-specificity profile to previously described hydrolases. Native diffraction data sets were collected from crystals of NH from each organism. The hexagonal crystals (space group P6(2)22 or P6(4)22) of NH from C. elegans diffracted to a resolution of 2.8 A, while the data set from the orthorhombic crystals (space group I222 or I2(1)2(1)2(1)) of NH from C. jejuni could be processed to 1.7 A resolution. The unit-cell parameters were a = b = 102.23, c = 117.27 A in the former case and a = 101.13, b = 100.13, c = 81.37 A in the latter.

Alternate JournalActa Crystallogr. D Biol. Crystallogr.
PubMed ID12777783