Phosphoglucomutase is absent in Trypanosoma brucei and redundantly substituted by phosphomannomutase and phospho-N-acetylglucosamine mutase. / Bandini, Giulia; Marino, Karina; Güther, M. Lucia Sampaio; Wernimont, Amy K.; Kuettel, Sabine; Qiu, Wei; Afzal, Shamshad; Kelner, Anna; Hui, Raymond; Ferguson, Michael A. J.
In: Molecular Microbiology, Vol. 85, No. 3, 2012, p. 513-534.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Phosphoglucomutase is absent in Trypanosoma brucei and redundantly substituted by phosphomannomutase and phospho-N-acetylglucosamine mutase
A1 - Bandini,Giulia
A1 - Marino,Karina
A1 - Güther,M. Lucia Sampaio
A1 - Wernimont,Amy K.
A1 - Kuettel,Sabine
A1 - Qiu,Wei
A1 - Afzal,Shamshad
A1 - Kelner,Anna
A1 - Hui,Raymond
A1 - Ferguson,Michael A. J.
AU - Bandini,Giulia
AU - Marino,Karina
AU - Güther,M. Lucia Sampaio
AU - Wernimont,Amy K.
AU - Kuettel,Sabine
AU - Qiu,Wei
AU - Afzal,Shamshad
AU - Kelner,Anna
AU - Hui,Raymond
AU - Ferguson,Michael A. J.
PY - 2012
Y1 - 2012
N2 - 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 1.85Å 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. © 2012 Blackwell Publishing Ltd.
AB - 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 1.85Å 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. © 2012 Blackwell Publishing Ltd.
UR - http://www.scopus.com/inward/record.url?partnerID=yv4JPVwI&eid=2-s2.0-84864109063&md5=4928c1298b086a5d87dbc9a807a9e3ee
U2 - 10.1111/j.1365-2958.2012.08124.x
DO - 10.1111/j.1365-2958.2012.08124.x
M1 - Article
JO - Molecular Microbiology
JF - Molecular Microbiology
SN - 0950-382X
IS - 3
VL - 85
SP - 513
EP - 534
ER -