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Volume 148, Issue 4

MabA (FabG1), a protein involved in the long-chain fatty acid elongation system FAS-II Free

Abstract

The fatty acid elongation system FAS-II is involved in the biosynthesis of mycolic acids, which are very long-chain fatty acids of the cell envelope specific to and other mycobacteria. A potential component of FAS-II, the protein MabA (FabG1), was overexpressed and purified. Sedimentation equilibrium analyses revealed that MabA undergoes a dimer to tetramer self-association with a dissociation constant of 22 μM. The protein was detected by Western blotting in a mycobacterial cell-wall extract that produces mycolic acids and in the FPLC FAS-II fraction. MabA was shown to catalyse the NADPH-specific reduction of β-ketoacyl derivatives, equivalent to the second step of a FAS-II elongation round. Unlike the known homologous proteins, MabA preferentially metabolizes long-chain substrates (C–C) and has a poor affinity for the C substrate, in agreement with FAS-II specificities. Molecular modelling of MabA structure suggested the presence of an unusually hydrophobic substrate-binding pocket holding a unique Trp residue, suitable for fluorescence spectroscopic analyses. In agreement with the enzyme kinetic data, the spectral properties of MabA were different in the presence of the C–C ligands as compared to the C ligand. Altogether, these data bring out distinctive enzymic and structural properties of MabA, which correlate with its predilection for long-chain substrates, in contrast to most of the other known ketoacyl reductases.

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Keyword(s): ACP, acyl carrier protein , enzymic activity , ESI, electrospray ionization , FAS, fatty acid synthetase , fluorescence spectroscopy , H-MabA, His-tagged MabA , INH, isoniazid , KAR, β-ketoacyl-ACP reductase , MALDI-TOF, matrix-associated laser desorption ionization-time of flight , PDB, Protein Data Bank , quaternary structure , SDR, short-chain dehydrogenases/reductases , structural model and β-ketoacyl reductase
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2002-04-01
2023-06-08
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MabA (FabG1), a Mycobacterium tuberculosis protein involved in the long-chain fatty acid elongation system FAS-II
Microbiology 148, 951 (2002); https://doi.org/10.1099/00221287-148-4-951
/content/journal/micro/10.1099/00221287-148-4-951
/content/journal/micro/10.1099/00221287-148-4-951
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