Molecular Basis of the Functional Divergence of Fatty Acyl-AMP Ligase Biosynthetic Enzymes of Mycobacterium tuberculosis

Mechanism and Regulation of Mycobactin Fatty Acyl-AMP Ligase FadD33

“…However, the activity of the FadD33-K511A mutant is completely abolished. It was shown in the FadD13 structure that the equivalent residue to FadD33 Lys-511, FadD13 Lys-487 faces the catalytic pocket in the acyl-adenylate-forming conformation (27) and that the FadD13-K487A mutant exhibits a 30-fold reduction of the k cat (28). Acetylation on Lys-511 is therefore likely to have functional consequences on FadD33 activity.…”

Insight into Structure-Function Relationships and Inhibition of the Fatty Acyl-AMP Ligase (FadD32) Orthologs from Mycobacteria

“…6 and Table 4). These structures included that of the N-terminal domain of the FAAL FadD28 (41) and those corresponding to the full-length FACL enzyme FadD13 and its N terminus (42,43). As the orientations of the N-and C-terminal domains differ between these two proteins, the superimposition of FadD13 and MmFadD32 structures was based on the use of N-or C-terminal domains.…”

Structures of Mycobacterium tuberculosis FadD10 Protein Reveal a New Type of Adenylate-forming Enzyme

“…For example, firefly luciferase (38) as well as five different adenylation domains of NRPSs (39) were shown to synthesize luciferylCoA and aminoacyl-CoAs, respectively, when their genuine acyl acceptors were absent, and coenzyme A was added as substrate. In contrast, M. tuberculosis FAALs (acyl-ACP synthetases) that have been studied to date, which have also been proposed to be descendants of acyl-CoA synthetases, lack acylCoA synthesis activity (3,13), even though they retain the binding elements of phosphopantetheine. This was demonstrated as well for FadD10.…”

“…However, it prevents the FAALs from undergoing the large scale inter-domain rotation associated with coenzyme A binding. The functional role of the insertion was further established by showing that its deletion and addition manipulated the gain and loss of acyl-CoA synthesis activity in M. tuberculosis FAALs and FACLs, respectively (3,13).…”

“…Recent structural and biochemical studies of M. tuberculosis FAAL28 (3) and FAALs from Escherichia coli and Legionella pneumophila (4) have provided a molecular basis for their catalytic mechanism. These studies have identified a signature sequence motif, consisting of an insertion of about 20 amino acids, that defines the function of this newly recognized subclass of the adenylate-forming superfamily and distinguishes them from FACLs (3,13).…”