Decrypting the oscillating nature of the 4'-phosphopantetheine arm in acyl carrier protein AcpM of Mycobacterium tuberculosis
Biswas R, Singh BK, Dutta D, Das PK, Maiti MK, Basak A, Das AK
FEBS letters · 2019-03
Abstract
In Mycobacterium tuberculosis, acyl carrier protein (AcpM)-mediated fatty acid synthase type II is integral for the synthesis of mycolic acids. AcpM, designated as an atypical ACP, comprises of a putative 33 amino acid long C-terminal extension which is distinctive in nature. Here, we aimed at devising an 'easy-to-go' method for the generation of crypto-AcpM loaded with a solvatochromic probe 7-Nitrobenz-2-oxa-1,3-diazol-4-yl, which is linked to the 4'-phosphopantetheine (Ppant) prosthetic group of AcpM. The crypto-AcpM, coupled with fluorescence spectroscopy and molecular dynamics simulation studies, was employed to explore the elusive dynamics of Ppant arm in AcpM. This investigation establishes the role of the flexible C-terminal extension of AcpM in regulating the prosthetic group sequestration ability by modulating the 'Asp-Ser-Leu' motif.
MeSH terms
- Escherichia coli
- Mycobacterium tuberculosis
- Mycolic Acids
- Nitrobenzenes
- Pantetheine
- Azoles
- Coenzyme A
- Bacterial Proteins
- Carrier Proteins
- Recombinant Proteins
- Fluorescent Dyes
- Cloning, Molecular
- Sequence Alignment
- Gene Expression
- Binding Sites
- Amino Acid Motifs
- Protein Binding
- Sequence Homology, Amino Acid
- Substrate Specificity
- Genetic Vectors
- Protein Interaction Domains and Motifs
- Molecular Dynamics Simulation
- Molecular Docking Simulation
- Protein Conformation, alpha-Helical