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Integrated multi-modal approach for structure and function determination of lignin-forming protein complexes isolated from plants


EMSL Project ID
51696

Abstract

We propose to apply an integrated multi-modal structural biology approach to investigate a biosynthetic protein complex in plants that is hypothesized to organize and guide lignin biosynthesis via oxidative coupling of propenylphenol precursors. This approach will apply multiple structural biology tools to the problem, including NMR to map substrate interactions, cross-linking MS and native MS to identify complex components and their positions in the complex, CryoEM to determine overall complex architecture, and computational modeling of the complex to tie data from different experimental studies together. Additional data obtained through an existing collaboration in X-ray diffraction (XRD) could also be incorporated. Integrated approaches are an emerging paradigm for understanding structure and function of protein complexes and will be applicable to other challenging systems of interest to EMSL users. This project will demonstrate such an approach on a difficult system and identify bottlenecks and unanticipated challenges in sample preparation and manipulation that would typically be encountered when working with EMSL users and their own systems of interest.

Project Details

Start Date
2020-12-15
End Date
2023-09-30
Status
Closed

Team

Principal Investigator

John Cort
Institution
Pacific Northwest National Laboratory

Co-Investigator(s)

Jay Bardhan
Institution
Environmental Molecular Sciences Laboratory

Team Members

Syed Moinuddin
Institution
Washington State University

Mischelle Schutz
Institution
Pacific Northwest National Laboratory

Madelyn Berger
Institution
Pacific Northwest National Laboratory

Stephanie Bobadilla-Regalado
Institution
Pacific Northwest National Laboratory

Briana Sobecks
Institution
Pacific Northwest National Laboratory

Margaret Cheung-Wyker
Institution
Environmental Molecular Sciences Laboratory

John Melchior
Institution
Pacific Northwest National Laboratory

Robert Young
Institution
Environmental Molecular Sciences Laboratory

Mowei Zhou
Institution
Environmental Molecular Sciences Laboratory

James Evans
Institution
Environmental Molecular Sciences Laboratory

Eric Merkley
Institution
Pacific Northwest National Laboratory