Access the full text.
Sign up today, get DeepDyve free for 14 days.
Loading next page...
/lp/de-gruyter/expression-and-characterization-of-recombinant-drosophila-6-pyruvoyl-0bdJcimIH3
References
References for this paper are not available at this time. We will be adding them shortly, thank you for your patience.
- Publisher
- de Gruyter
- Copyright
- Copyright © 1995 by the
- ISSN
- 0933-4807
- eISSN
- 2195-4720
- DOI
- 10.1515/pteridines.1995.6.2.58
- Publisher site
- See Article on Publisher Site
Abstract
Summary 6-Pyruvoyl tetrahydropterin synthase is involved in the synthesis of pteridine eye pigments in Drosophila. The purple gene which was known to be one of the target loci of the suppressor mutation su(sj2 has been identified to encode the enzyme, and its cDNA has been cloned recently. The cDNA encoding the 19.3 kDa subunit of the 6-pyruvoyl tetrahydropterin synthase was expressed as fusion proteins in E. coli. The recombinant protein was shown to be active and purified from the E. coli crude extract by metal-chelation chromatography. The fused metal-chelating oilgopeptide was removed by thrombin for further characterization. Apparent Km for the substrate dihydroneopterin triphosphate was determined to be 590 IlM, which was slightly higher than the value of the native enzyme. The isoelectric point of 6.4 was also different from the known value of 4.3 determined by the native enzyme. Heat stability and the stimulatory effect of reducing agents were similar to the native enzyme. The modification of cysteine residues in the recombinant enzyme, one of which is known to be conserved in human and rat enzymes, by iodoacetamide inhibited its activity by up to 80%.
Journal
Pteridines – de Gruyter
Published: May 1, 1995