Crystal structures of a putative periplasmic cystine-binding protein from Candidatus Liberibacter asiaticus: insights into an adapted mechanism of ligand binding.

ABC transporters Candidatus Liberibacter asiaticus crystal structure periplasmic amino acid-binding protein surface plasmon resonance

Journal

The FEBS journal
ISSN: 1742-4658
Titre abrégé: FEBS J
Pays: England
ID NLM: 101229646

Informations de publication

Date de publication:
09 2019
Historique:
received: 25 01 2019
revised: 20 03 2019
accepted: 03 05 2019
pubmed: 8 5 2019
medline: 27 5 2020
entrez: 8 5 2019
Statut: ppublish

Résumé

The amino acid-binding receptors, a component of ABC transporters, have evolved to cater to different specificities and functions. Of particular interest are cystine-binding receptors, which have shown broad specificity. In the present study, a putative periplasmic cystine-binding protein from Candidatus Liberibacter asiaticus (CLasTcyA) was characterized. Analysis of the CLasTcyA sequence and crystal structures in the ligand-bound state revealed novel features of CLasTcyA in comparison to related proteins. One of the unique features found in CLasTcyA structure was the positioning of the C-terminal extended loop of one chain very close to the substrate-binding site of the adjacent monomer in the asymmetric unit. The presence of a disulphide bond, unique to Candidatus Liberibacter family, holds the C-terminal extended loop in position. Analysis of the substrate-binding pocket of CLasTcyA suggested a broad specificity and a completely different orientation of the bound substrates in comparison to related protein structures. The open conformation for one of the two chains of the asymmetric unit in the Arg-bound structure revealed a limited open state (18.4°) for CLasTcyA as compared to open state of other related proteins (~ 60°). The strong interaction between Asp126 on helix-α5 of small domain and Arg82 (bigger domain) restricts the degree of opening in ligand-free open state. The dissociation constant of 1.26 μm by SPR and 3.7 μm by MST exhibited low affinity for the cystine. This is the first structural characterization of an l-cystine ABC transporter from plant pathogen and our results suggest that CLasTcyA may have evolved to cater to its specific needs for its survival in the host.

Identifiants

pubmed: 31063259
doi: 10.1111/febs.14921
doi:

Substances chimiques

Bacterial Proteins 0
Cysteine K848JZ4886

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

3450-3472

Subventions

Organisme : UGC, Government of India
Pays : International

Informations de copyright

© 2019 Federation of European Biochemical Societies.

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Auteurs

Pranav Kumar (P)

Department of Biotechnology, Indian Institute of Technology Roorkee, India.

Pooja Kesari (P)

Department of Biotechnology, Indian Institute of Technology Roorkee, India.

Sunil Kokane (S)

Plant Virology Laboratory, ICAR-Central Citrus Research Institute, Nagpur, India.

Dilip Kumar Ghosh (DK)

Plant Virology Laboratory, ICAR-Central Citrus Research Institute, Nagpur, India.

Pravindra Kumar (P)

Department of Biotechnology, Indian Institute of Technology Roorkee, India.

Ashwani Kumar Sharma (AK)

Department of Biotechnology, Indian Institute of Technology Roorkee, India.

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