Phosphoglucomutase is absent in Trypanosoma brucei and redundantly substituted by phosphomannomutase and phospho-N-acetylglucosamine mutase.
The enzymes phosphomannomutase (PMM), phospho-N-acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between the C6 and C1 hydroxyl groups of mannose, N-acetylglucosamine and glucose respectively. Although genes for a candidate PMM and a PAGM enzymes have been found in the Trypanosoma brucei genome, there is, surprisingly, no candidate gene for PGM. The TbPMM and TbPAGM genes were cloned and expressed in Escherichia coli and the TbPMM enzyme was crystallized and its structure solved at 1.85 Å resolution. Antibodies to the recombinant proteins localized endogenous TbPMM to glycosomes in the bloodstream form of the parasite, while TbPAGM localized to both the cytosol and glycosomes. Both recombinant enzymes were able to interconvert glucose-phosphates, as well as acting on their own definitive substrates. Analysis of sugar nucleotide levels in parasites with TbPMM or TbPAGM knocked down by RNA interference (RNAi) suggests that, in vivo, PGM activity is catalysed by both enzymes. This is the first example in any organism of PGM activity being completely replaced in this way and it explains why, uniquely, T. brucei has been able to lose its PGM gene. The RNAi data for TbPMM also showed that this is an essential gene for parasite growth.
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
Giulia
G
Bandini
Karina
K
Mariño
M Lucia Sampaio
ML
Güther
Amy K
AK
Wernimont
Sabine
S
Kuettel
Wei
W
Qiu
Shamshad
S
Afzal
Anna
A
Kelner
Raymond
R
Hui
Michael A J
MA
Ferguson
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019298:Glucose-6-Phosphate / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D007700:Kinetics; D008360:Mannosephosphates / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010733:Phosphoglucomutase / Q000172:deficiency*; D017875:Phosphotransferases (Phosphomutases) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D021381:Protein Transport; D034622:RNA Interference; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics*
D005958:Glucosephosphates; D008360:Mannosephosphates; D011994:Recombinant Proteins; D019298:Glucose-6-Phosphate; C026146:N-acetylglucosamine-1-phosphate; C031590:glucose-1-phosphate; D017875:Phosphotransferases (Phosphomutases); D010733:Phosphoglucomutase; C052799:phosphomannomutase; D000117:Acetylglucosamine