Cloning, sequencing and expression of ribonucleotide reductase R2 from Trypanosoma brucei

• Published in: FEBS letters Vol. 414; no. 2; pp. 449 - 453
• Main Authors: Dormeyer, Matthias, Schöneck, Ralf, Dittmar, Gunnar A.G., Krauth-Siegel, R. Luise
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 08.09.1997
• Subjects: Amino Acid Sequence; Animals; Cloning, Molecular; DNA, Complementary; DNA, Protozoan - chemistry; Drug target; Escherichia coli; Escherichia coli - enzymology; Genes, Protozoan; Humans; Mice; Molecular Sequence Data; Plasmodium falciparum - enzymology; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Ribonucleotide reductase; Ribonucleotide Reductases - biosynthesis; Ribonucleotide Reductases - chemistry; Ribonucleotide Reductases - genetics; Sequence Alignment; Sequence Homology, Amino Acid; Trypanosoma brucei; Trypanosoma brucei brucei - enzymology; Trypanosoma brucei brucei - genetics; Trypanothione; RR; R2; nrd B; Ribonucleotide reductase; Drug target; IPTG; T.b; Trypanosoma brucei; Trypanothione; Ni-NTA; R1
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)01036-3

Ribonucleotide reductase (RR) is an attractive drug target molecule. The gene of the R2 protein of Trypanosoma brucei RR ( nrd B) has been cloned. It encodes a protein of 337 residues which shows about 60% identity with other eukaryotic R2 proteins. All residues which bind the iron center, the tyrosyl radical or are supposed to participate in the radical transfer are conserved in the trypanosomal protein sequence. Overexpression of the gene in E. coli resulted in 2–5 mg pure R2 protein from 100 ml bacterial cell culture. Northern blot analysis revealed a transcript of 1.85 kb in bloodstream and procyclic forms of the parasite.