An essential GTPase, der, containing double GTP-binding domains from Escherichia coli and Thermotoga maritima.
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Hwang J, Inouye M
An essential GTPase, der, containing double GTP-binding domains from Escherichia coli and Thermotoga maritima.
J Biol Chem. 2001 Aug 17;276(33):31415-21. Epub 2001 May 31.
- PubMed ID
- 11387344 [ View in PubMed]
- Abstract
A gene encoding a putative GTPase containing two tandemly repeated GTP-binding domains from a hyperthermophilic bacterium, Thermotoga maritima, was cloned and expressed in Escherichia coli. The gene (TM1446) termed der is highly conserved in Eubacteria including E. coli. The purified der product (Tm-Der) has GTPase activity but no ATPase activity. GTP, GDP, and dGTP but not GMP, ATP, CTP, and UTP compete for GTP binding to Tm-Der. An optimal condition for the GTPase assay was determined to be pH 7.5 in 400 mm KCl and 5 mm MgCl(2) at 70 degrees C, where K(m), V(max), and k(cat) values were determined to be 110 microm, 3.46 microm/min, and 0.87 min(-1), respectively. A der deletion strain of E. coli was constructed by replacing the der gene (originally annotated yfgK) with a kanamycin resistance gene. The deletion strain was found to form colonies only if the cells harbored a plasmid containing der, indicating that der is essential for E. coli growth.