TY - JOUR
T1 - Allosteric pyruvate kinase-based "logic gate" synergistically senses energy and sugar levels in Mycobacterium tuberculosis
AU - Zhong, Wenhe
AU - Cui, Liang
AU - Goh, Boon Chong
AU - Cai, Qixu
AU - Ho, Peiying
AU - Chionh, Yok Hian
AU - Yuan, Meng
AU - Sahili, Abbas El
AU - Fothergill-Gilmore, Linda A
AU - Walkinshaw, Malcolm D
AU - Lescar, Julien
AU - Dedon, Peter C
PY - 2017/12/7
Y1 - 2017/12/7
N2 - Pyruvate kinase (PYK) is an essential glycolytic enzyme that controls glycolytic flux and is critical for ATP production in all organisms, with tight regulation by multiple metabolites. Yet the allosteric mechanisms governing PYK activity in bacterial pathogens are poorly understood. Here we report biochemical, structural and metabolomic evidence that Mycobacterium tuberculosis (Mtb) PYK uses AMP and glucose-6-phosphate (G6P) as synergistic allosteric activators that function as a molecular "OR logic gate" to tightly regulate energy and glucose metabolism. G6P was found to bind to a previously unknown site adjacent to the canonical site for AMP. Kinetic data and structural network analysis further show that AMP and G6P work synergistically as allosteric activators. Importantly, metabolome profiling in the Mtb surrogate, Mycobacterium bovis BCG, reveals significant changes in AMP and G6P levels during nutrient deprivation, which provides insights into how a PYK OR gate would function during the stress of Mtb infection.
AB - Pyruvate kinase (PYK) is an essential glycolytic enzyme that controls glycolytic flux and is critical for ATP production in all organisms, with tight regulation by multiple metabolites. Yet the allosteric mechanisms governing PYK activity in bacterial pathogens are poorly understood. Here we report biochemical, structural and metabolomic evidence that Mycobacterium tuberculosis (Mtb) PYK uses AMP and glucose-6-phosphate (G6P) as synergistic allosteric activators that function as a molecular "OR logic gate" to tightly regulate energy and glucose metabolism. G6P was found to bind to a previously unknown site adjacent to the canonical site for AMP. Kinetic data and structural network analysis further show that AMP and G6P work synergistically as allosteric activators. Importantly, metabolome profiling in the Mtb surrogate, Mycobacterium bovis BCG, reveals significant changes in AMP and G6P levels during nutrient deprivation, which provides insights into how a PYK OR gate would function during the stress of Mtb infection.
KW - Enzyme mechanisms
KW - Infectious diseases
KW - Metabolomics
KW - Pathogens
KW - X-ray crystallography
UR - https://www.rcsb.org/structure/5WRP
UR - https://www.rcsb.org/structure/5WS8
UR - https://www.rcsb.org/structure/5WS9
UR - https://www.rcsb.org/structure/5WSA
UR - https://www.rcsb.org/structure/5WSB
UR - https://www.rcsb.org/structure/5WSC
U2 - 10.1038/s41467-017-02086-y
DO - 10.1038/s41467-017-02086-y
M3 - Article
C2 - 29215013
SN - 2041-1723
VL - 8
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 1986
ER -