Molecular cloning and expression of a purine specific N-ribohydrolase from trypanosoma bruce1 brvcel

R. Pelle, Vern L. Schramm, D. W. Parkin

Research output: Contribution to journalArticle

Abstract

N-Ribohydrolases, including the inosine-adenosine-guanosine preferring nucleoside hydrolase (IAG-NH), are involved in the nucleoside salvage pathway of African trypanosomes. The iagnh gene contains an open reading frame (ORF) of 981 bp, is present as one copy/haploid genome, and is located on the size-polymorphic chromosome III or IV in the genome of '/'. h. hrncei On Southern analysis the IAG-NH probe hybridizes strongly with /'. conglense and /'. vivax and to a lesser extent with T. cruzi and l.inshmania thmovani genomic DNA. The iagnh gene is expressed in all the life-cycle-stages of T. b. brucei. There is no known significant amino acid identity of the IAG-NH sequence outside bacteria, yeast, or parasitic organisms, with only low amino acid homology occuring with the inosineuridine preferring (IU-) NH isozyme from (". fascicnlata. The iagnh (ORF) was cloned into K. coli BL21(DE3) and a soluble recombinant IAG-NH expressed and purified to > 97% homogeneity. The molecular weight of the recombinant IAG-NH, based on the amino acid sequence and observed mass, was 35.735 and 35,737, respectively. Conserved motifs DxDxxxDDxxA and AExNI have been identified in N-ribohydrolases and are associated with Ca and ribose binding.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997

Fingerprint

Trypanosoma
Cloning
Molecular Cloning
purines
molecular cloning
Genes
Amino Acids
Open Reading Frames
open reading frames
Genome
Salvaging
amino acids
Ribose
genome
Trypanosomiasis
ribose
nucleosides
Haploidy
Chromosomes
Life Cycle Stages

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Molecular cloning and expression of a purine specific N-ribohydrolase from trypanosoma bruce1 brvcel. / Pelle, R.; Schramm, Vern L.; Parkin, D. W.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

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