Abstract
The unique and ubiquitous bacterial lipoprotein biosynthesis pathway is an attractive new antibiotic target. Crystal structures of its three biosynthetic enzymes have been solved recently. The first enzyme, Phosphatidylglycerol:proLipoprotein diacylglyceryl Transferase (Lgt), which initiates the post-translational modification at the metabolic interface of protein biosynthesis, phospholipid biosynthesis, protein secretion and lipid modification was reported to be a seven-transmembrane helical structure with a catalytic periplasmic head. Its complete solubilization in water or mild detergent in a fully active state, its chromatographic behaviour as an active monomer in the absence of detergent and recovery of active whole-length protein after proteolytic treatment of spheroplasts cast serious doubts about its proposed membrane association and orientation. Rather, it could be a seven-helical bundle partially embedded in the inner membrane’s inner leaflet aided by hydrophobic interaction. In fact, there are examples where originally reported seven-transmembrane proteins were later shown to be seven-helical peripheral membrane proteins based on solubilization criterion and re-analysis. Validated computational tool, Membrane Optimal Docking Area (MODA), also predicted a weaker association of Lgt’s helices with the membrane compared to typical transmembrane proteins. This insight is crucial to Lgt-based antibiotic design.
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Abbreviations
- Lgt:
-
Phosphatidylglycerol:proLipoprotein diacylglyceryl transferase
- LspA:
-
Lipoprotein signal peptidase II
- Lnt:
-
apoLipoprotein N-acyl transferase
- GPCR:
-
G protein-coupled receptor
- MODA:
-
Membrane Optimal Docking Area
- VADAR:
-
Volume Area Dihedral Angle Reporter
- PG:
-
Phosphatidylglycerol
- IMV:
-
Inverted membrane vesicles
- OG:
-
n-octyl-β-d-glucoside
- ASA:
-
Accessible surface area
- PDB:
-
Protein Data Bank
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Acknowledgement
We acknowledge the support from DBT-BUILDER Programme (BT/PR12153/INF/22/200/2014), Department of Biotechnology, Government of India, New Delhi. We gratefully acknowledge the critical reading, comments and inputs of Dr. M. Madan Babu of LMB, Cambridge, UK.
Funding
This study was funded by the Department of Biotechnology, BUILDER programme, Government of India (BT/PR12153/INF/22/200/2014).
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NS performed cloning, expression and Lgt fluorescent assay; conceived and performed the in silico analysis and wrote the manuscript. SK performed solubilization and gel mobility assays. KS conceived the concept, mentored, wrote and edited the manuscript.
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Sangith, N., Kumar, S. & Sankaran, K. Evidence to Suggest Bacterial Lipoprotein Diacylglyceryl Transferase (Lgt) is a Weakly Associated Inner Membrane Protein. J Membrane Biol 252, 563–575 (2019). https://doi.org/10.1007/s00232-019-00076-3
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DOI: https://doi.org/10.1007/s00232-019-00076-3