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Structural Characterization of Acidic M17 Leucine Aminopeptidases from the TriTryps and Evaluation of Their Role in Nutrient Starvation in Trypanosoma brucei

DOI: 10.1128/mSphere.00226-17 DOI Help

Authors: Jennifer Timm (University of York) , Maria Valente (Parasitología y Biomedicina López-Neyra, Consejo Superior de Investigaciones Científicas) , Daniel García-caballero (Parasitología y Biomedicina López-Neyra, Consejo Superior de Investigaciones Científicas) , Keith Wilson (University of York) , Dolores González-pacanowska (Parasitología y Biomedicina López-Neyra, Consejo Superior de Investigaciones Científicas)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Msphere , VOL 2

State: Published (Approved)
Published: August 2017
Diamond Proposal Number(s): 7864 , 9948

Abstract: Leucine aminopeptidase (LAP) is found in all kingdoms of life and catalyzes the metal-dependent hydrolysis of the N-terminal amino acid residue of peptide or amino acyl substrates. LAPs have been shown to participate in the N-terminal processing of certain proteins in mammalian cells and in homologous recombination and transcription regulation in bacteria, while in parasites, they are involved in host cell invasion and provision of essential amino acids for growth. The enzyme is essential for survival in Plasmodium falciparum, where its drug target potential has been suggested. We report here the X-ray structures of three kinetoplastid acidic LAPs (LAP-As from Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major) which were solved in the metal-free and unliganded forms, as well as in a number of ligand complexes, providing insight into ligand binding, metal ion requirements, and oligomeric state. In addition, we analyzed mutant cells defective in LAP-A in Trypanosoma brucei, strongly suggesting that the enzyme is not required for the growth of this parasite either in vitro or in vivo. In procyclic cells, LAP-A was equally distributed throughout the cytoplasm, yet upon starvation, it relocalizes in particles that concentrate in the perinuclear region. Overexpression of the enzyme conferred a growth advantage when parasites were grown in leucine-deficient medium. Overall, the results suggest that in T. brucei, LAP-A may participate in protein degradation associated with nutrient depletion.

Journal Keywords: Leishmania major; Trypanosoma brucei; Trypanosoma cruzi; crystallography; knockout; leucine aminopeptidase

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength) , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography