Structural and Binding Properties of the Active Cell Wall Hydrolase RipA from M. tuberculosis, a Promising Biosensing Molecule for Early Warning Bacterial Detection
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Structural and Binding Properties of the Active Cell Wall Hydrolase RipA from M. tuberculosis, a Promising Biosensing Molecule for Early Warning Bacterial Detection (126 views)

Squeglia F, Marasco D, Ruggiero A, Testa G, Esposito L, Berisio R

Curr Med Chem (ISSN: 0929-8673linking, 1875-533xelectronic), 2022; 29(24): 4282-4292.

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Paper type: Journal Article,

Impact factor: 4.74, 5-year impact factor: 4.74

Url: Not available.

Keywords: Bacterial Proteins Metabolism, Cell Wall Metabolism, Endopeptidases Metabolism, Hydrolases Metabolism, Mycobacterium Tuberculosis Metabolism, Peptidoglycan Metabolism, Binding, Structure,

Affiliations:
*** IBB - CNR ***

Institute of Biostructures and Bioimaging, IBB, CNR, Naples, Italy.
Department of Pharmacy, University of Naples, Naples, Italy.
Institute for electromagnetic sensing of the environment, IREA, CNR, Naples, Italy.


References:
Not available.
Structural and Binding Properties of the Active Cell Wall Hydrolase RipA from M. tuberculosis, a Promising Biosensing Molecule for Early Warning Bacterial Detection

BACKGROUND: Peptidoglycan is an essential component of the cell wall in all bacteria. In particular, the cell walls of Gram-positive bacteria are composed mostly of a thick layer of peptidoglycan. Its accessibility has important implications for their sensing in whole bacterial detection methodologies. Indeed, there is an urgent demand for rapid tests which can identify whole bacteria, e.g., directly at the point of care. OBJECTIVE: The aim of this work is to explore the suitability of RipA, a key cell division protein of M. tuberculosis, for whole cell biosensing of Gram-positive bacteria. METHODS: We here conducted Molecular Dynamics (MD) studies aimed at the understanding of the structural and dynamic features of active RipA and at the design of a suitable bioreceptor. Based on these studies, we engineered a RipA variant for covalent oriented immobilisation on golden surfaces and are able to bind peptidoglycan, albeit without degrading it. Surface Plasmon Resonance (SPR) was employed to check the ability of functionalized golden chips to recognize whole bacteria. RESULTS: MD analyses elucidated the structural details of the active form of RipA and suggested that this enzyme, once inactivated, presents a rigid and well-exposed peptidoglycan recognition cleft. We engineered RipA for proper oriented immobilisation on golden chips for SPR studies. Results show that once chemically coupled to a golden chip, the developed RipA-based bioreceptor is able to detect B. subtilis, used as a model in a concentration-dependent mode. CONCLUSION: Results highlight the potential of the engineered molecule to be integrated in the development of early warning biosensors for Gram-positive contamination in clinical diagnosis or food-borne infections.
Structural and Binding Properties of the Active Cell Wall Hydrolase RipA from M. tuberculosis, a Promising Biosensing Molecule for Early Warning Bacterial Detection

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Structural and Binding Properties of the Active Cell Wall Hydrolase RipA from M. tuberculosis, a Promising Biosensing Molecule for Early Warning Bacterial Detection

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Squeglia F, Moreira M, Ruggiero A, Berisio R
* The Cell Wall Hydrolytic NlpC/P60 Endopeptidases in Mycobacterial Cytokinesis: A Structural Perspective (390 views)
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Squeglia F, Ruggiero A, Berisio R
* Collagen degradation in tuberculosis pathogenesis: the biochemical consequences of hosting an undesired guest (566 views)
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Squeglia F, Ruggiero A, Berisio R
* Chemistry of Peptidoglycan in Mycobacterium tuberculosis Life Cycle: An off-the-wall Balance of Synthesis and Degradation (255 views)
Chemistry (ISSN: 0947-6539linking, 1521-3765electronic), 2018 Feb 21; 24 (11 ): 2533-2546.
Impact Factor: 5.476
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Ruggiero A, Squeglia F, Romano M, Vitagliano L, De Simone A, Berisio R
* Structure and dynamics of the multi-domain resuscitation promoting factor RpfB from Mycobacterium tuberculosis (536 views)
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Palmieri G, Cocca E, Gogliettino M, Valentino R, Ruvo M, Cristofano G, Angiolillo A, Balestrieri M, Rossi M, Di Costanzo A
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Esposito C, Cantisani M, D'Auria G, Falcigno L, Pedone E, Galdiero S, Berisio R
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12 Records (12 excluding Abstracts).
Total impact factor: 45.41 (45.41 excluding Abstracts).
Total 5 year impact factor: 40.279 (40.279 excluding Abstracts).

Last modified by Rita Berisio on Tuesday 27 December 2022, 19:17:30
126 views. Last view on Sunday 01 December 2024, 5:00:33