title: The trifunctional sulfate-activating complex (SAC) of Mycobacterium tuberculosis.
abstract: The sulfate activation pathway is essential for the assimilation of sulfate and, in many bacteria, is comprised of three reactions: the synthesis of adenosine 5'-phosphosulfate (APS), the hydrolysis of GTP, and the 3'-phosphorylation of APS to produce 3'-phosphoadenosine 5'-phosphosulfate (PAPS), whose sulfuryl group is reduced or transferred to other metabolites. The entire sulfate activation pathway is organized into a single complex in Mycobacterium tuberculosis. Although present in many bacteria, these tripartite complexes have not been studied in detail. Initial rate characterization of the mycobacterial system reveals that it is poised for extremely efficient throughput: at saturating ATP, PAPS synthesis is 5800 times more efficient than APS synthesis. The APS kinase domain of the complex does not appear to form the covalent E.P intermediate observed in the closely related APS kinase from Escherichia coli. The stoichiometry of GTP hydrolysis and APS synthesis is 1:1, and the APS synthesis reaction is driven 1.1 x 10(6)-fold further during GTP hydrolysis; the system harnesses the full chemical potential of the hydrolysis reaction to the synthesis of APS. A key energy-coupling step in the mechanism is a ligand-induced isomerization that enhances the affinity of GTP and commits APS synthesis and GTP hydrolysis to the completion of the catalytic cycle. Ligand-induced increases in guanine nucleotide affinity observed in the mycobacterial system suggest that it too undergoes the energy-coupling isomerization.
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 10461-1926, USA.
authors.forename: Meihao
authors.initials: M
authors.lastname: Sun
authors.forename: John L
authors.initials: JL
authors.lastname: Andreassi
authors.forename: Shuqing
authors.initials: S
authors.lastname: Liu
authors.forename: Rachel
authors.initials: R
authors.lastname: Pinto
authors.forename: James A
authors.initials: JA
authors.lastname: Triccas
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Leyh
mesh_terms: D000250:Adenosine Phosphosulfate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D002384:Catalysis; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D006147:Guanine / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013430:Sulfate Adenylyltransferase / Q000737:chemistry; D013431:Sulfates / Q000737:chemistry*; D013997:Time Factors
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D013431:Sulfates; D000250:Adenosine Phosphosulfate; D006147:Guanine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D013430:Sulfate Adenylyltransferase