Dataset Information

CryoEM Structure of the, UND-PP bound, WaaL O-Antigen Ligase

ABSTRACT:

SUBMITTER: Khuram Ashraf

PROVIDER: EMPIAR-10938 | biostudies-other |

REPOSITORIES: biostudies-other

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Publications

Structural basis of lipopolysaccharide maturation by the O-antigen ligase.

Ashraf Khuram U KU Nygaard Rie R Vickery Owen N ON Erramilli Satchal K SK Herrera Carmen M CM McConville Thomas H TH Petrou Vasileios I VI Giacometti Sabrina I SI Dufrisne Meagan Belcher MB Nosol Kamil K Zinkle Allen P AP Graham Chris L B CLB Loukeris Michael M Kloss Brian B Skorupinska-Tudek Karolina K Swiezewska Ewa E Roper David I DI Clarke Oliver B OB Uhlemann Anne-Catrin AC Kossiakoff Anthony A AA Trent M Stephen MS Stansfeld Phillip J PJ Mancia Filippo F

Nature 20220406 7905

The outer membrane of Gram-negative bacteria has an external leaflet that is largely composed of lipopolysaccharide, which provides a selective permeation barrier, particularly against antimicrobials<sup>1</sup>. The final and crucial step in the biosynthesis of lipopolysaccharide is the addition of a species-dependent O-antigen to the lipid A core oligosaccharide, which is catalysed by the O-antigen ligase WaaL<sup>2</sup>. Here we present structures of WaaL from Cupriavidus metallidurans, both ...[more]

PMID: 35388216

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Project description:Adenoviral (Ad) vectors show promise as platforms for vaccine applications against infectious diseases including HIV. However, the requirements for eliciting protective neutralizing antibody and cellular immune responses against HIV remain a major challenge. In a novel approach to generate 2F5- and 4E10-like antibodies, we engineered an Ad vector with the HIV membrane proximal ectodomain region (MPER) epitope displayed on the hypervariable region 2 (HVR2) of the viral hexon capsid, instead of expressed as a transgene. The structure and flexibility of MPER epitopes, and the structural context of these epitopes within viral vectors, play important roles in the induced host immune responses. In this regard, understanding the critical factors for epitope presentation would facilitate optimization strategies for developing viral vaccine vectors. Therefore we undertook a cryoEM structural study of this Ad vector, which was previously shown to elicit MPER-specific humoral immune responses. A subnanometer resolution cryoEM structure was analyzed with guided molecular dynamics simulations. Due to the arrangement of hexons within the Ad capsid, there are twelve unique environments for the inserted peptide that lead to a variety of conformations for MPER, including individual α-helices, interacting α-helices, and partially extended forms. This finding is consistent with the known conformational flexibility of MPER. The presence of an extended form, or an induced extended form, is supported by interaction of this vector with the human HIV monoclonal antibody 2F5, which recognizes 14 extended amino acids within MPER. These results demonstrate that the Ad capsid influences epitope structure, flexibility and accessibility, all of which affect the host immune response. In summary, this cryoEM structural study provided a means to visualize an epitope presented on an engineered viral vector and suggested modifications for the next generation of Ad vectors with capsid-incorporated HIV epitopes.

Dataset Information

CryoEM Structure of the, UND-PP bound, WaaL O-Antigen Ligase

Publications

Structural basis of lipopolysaccharide maturation by the O-antigen ligase.

Similar Datasets

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