GDP-mannose pyrophosphorylase is essential in the bloodstream form of Trypanosoma brucei

The Biochemical Journal
Helen DentonTerry K Smith

Abstract

A putative GDP-Man PP (guanidine diphosphomannose pyrophosphorylase) gene from Trypanosoma brucei (TbGDP-Man PP) was identified in the genome and subsequently cloned, sequenced and recombinantly expressed, and shown to be a catalytically active dimer. Kinetic analysis revealed a Vmax of 0.34 mumol/min per mg of protein and Km values of 67 muM and 12 muM for GTP and mannose 1-phosphate respectively. Further kinetic studies showed GDP-Man was a potent product feedback inhibitor. RNAi (RNA interference) of the cytosolic TbGDP-Man PP showed that mRNA levels were reduced to ~20% of wild-type levels, causing the cells to die after 3-4 days, demonstrating that TbGDP-Man PP is essential in the bloodstream form of T. brucei and thus a potential drug target. The RNAi-induced parasites have a greatly reduced capability to form GDP-Man, leading ultimately to a reduction in their ability to synthesize their essential GPI (glycosylphosphatidylinositol) anchors. The RNAi-induced parasites also showed aberrant N-glycosylation of their major cell-surface glycoprotein, variant surface glycoprotein, with loss of the high-mannose Man9GlcNAc2 N-glycosylation at Asn428 and formation of complex N-glycans at Asn263.

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Citations

Apr 13, 2010·Molecular and Biochemical Parasitology·Terry K Smith, Peter Bütikofer
Jun 10, 2010·Organic & Biomolecular Chemistry·Sven NiewiadomskiGerd K Wagner
Jun 20, 2019·Frontiers in Cellular and Infection Microbiology·Sébastien PomelPhilippe M Loiseau
Jan 23, 2019·Frontiers in Cellular and Infection Microbiology·Juan Felipe Osorio-Méndez, Ana María Cevallos
Feb 17, 2021·PLoS Neglected Tropical Diseases·Maria Lucia Sampaio GutherMichael A J Ferguson

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African trypanosomiasis, also known as sleeping sickness, is an insect-borne parasitic disease of humans and other animals. It is caused by protozoa of the species Trypanosoma brucei and almost invariably progresses to death unless treated. Discover the latest research on African trypanosomiasis here.