Structural and Functional Studies of a Klebsiella Phage Capsule Depolymerase Tailspike: Mechanistic Insights into Capsular Degradation(39 views visite) Squeglia F, Maciejewska B, Łątka A, Ruggiero A, Briers Y, Drulis-kawa Z, Berisio R
Structure (ISSN: 0969-2126linking), 2020 Jun 2; 28(6): 613-624.
Keywords Parole chiave: Isothermal Titration Calorimetry
, Klebsiella Pneumoniae Capsule
, Crystal Structure
, Glycan Binding
, Infectious Disease
, Phage Depolymerase
, Protein
, Tailspike Branching System
Affiliations Affiliazioni: *** IBB - CNR ***
Institute of Biostructures and Bioimaging, CNR, Napoli, Italy.
Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, Poland.
Laboratory of Applied Biotechnology, Department of Biotechnology, Ghent University, Ghent, Belgium.
References Riferimenti: Not available. Non disponibili.
Structural and Functional Studies of a Klebsiella Phage Capsule Depolymerase Tailspike: Mechanistic Insights into Capsular Degradation
Capsule polysaccharide is a major virulence factor of Klebsiella pneumoniae, a nosocomial pathogen associated with a wide range of infections. It protects bacteria from harsh environmental conditions, immune system response, and phage infection. To access cell wall-located receptors, some phages possess tailspike depolymerases that degrade the capsular polysaccharide. Here, we present the crystal structure of a tailspike against Klebsiella, KP32gp38, whose primary sequence shares no similarity to other proteins of known structure. In the trimeric structure of KP32gp38, each chain contains a flexible N-terminal domain, a right-handed parallel β helix domain and two β sandwiches with carbohydrate binding features. The crystal structure and activity assays allowed us to locate the catalytic site. Also, our data provide experimental evidence of a branching architecture of depolymerases in KP32 Klebsiella viruses, as KP32gp38 displays nanomolar affinity to another depolymerase from the same phage, KP32gp37. Results provide a structural framework for enzyme engineering to produce serotype-broad-active enzyme complexes against K. pneumoniae.
Structural and Functional Studies of a Klebsiella Phage Capsule Depolymerase Tailspike: Mechanistic Insights into Capsular Degradation
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