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Structural Dynamics of Plant Cellulose Synthesis & Assembly Revealed by CryoTEM and DTEM


EMSL Project ID
48343

Abstract

We propose to capitalize on preliminary work defining in vitro cellulose synthesis from membrane protein purified from the nonvascular plant Physcomitrella patens. This system grants access to functionally active cellulose synthase under conditions that make cellulose fibrils. To exploit this opportunity to gain structural information about the plant machine that makes cellulose, and to visualize beta-1,4 glucan chains coalescing into fibers, we request access to EMSL's world class liquid helium-cooled cryoTEM, its novel dynamic TEM, and its outstanding computational capacity. These assets will be used to collect single particle images from TEM micrographs of both static and dynamically active cellulose synthase, and then process them with modern 3D reconstruction algorithms to determine the long sought structures. The DTEM is uniquely capable of providing the needed ultrafast imaging, and the robust computational capacities are essential for sorting through the heterogeneity expected for cellulose synthase in catalytic action. We anticipate a high impact paper on these findings - obtaining the requested EMSL support will make an even more convincing case and enhance the scope of the discovery. Achieving this cutting-edge research will significantly advance three important missions. First, the project offers a strong opportunity for EMSL to provide integrated experimental and computational resources for discovery to address complex energy and environmental issues facing the DOE and the Nation. In particular, it addresses the EMSL Science Research Theme of "Biosystem Dynamics and Design" - providing characterization of the machine that makes the key biological polymer from which biofuels can be produced from plant biomass and delivering structural information about a crucial transmembrane protein whose function dramatically impacts plant morphology and growth. Second, the planed experiments advance the mission of the DOE-EFRC Center for Lignocellulose Structure and Function which seeks to provide a detailed understanding of cellulose synthesis and fibril formation as a precursor to the biogenesis of plant cell walls. Finally, in all of its aspects the research advances toward the larger goal of the DOE to establish economical use of plant biomass as a renewable resource for production of biofuels.

Project Details

Project type
Large-Scale EMSL Research
Start Date
2014-10-01
End Date
2017-09-30
Status
Closed

Team

Principal Investigator

Bryan Nixon
Institution
Pennsylvania State University

Co-Investigator(s)

Susan Hafenstein
Institution
Pennsylvania State University

Team Members

Noopur Sharma
Institution
Environmental Molecular Sciences Laboratory

Sung Hyun Cho
Institution
Pennsylvania State University

Manish Kumar
Institution
Pennsylvania State University

Juan Du
Institution
Pennsylvania State University

Gong Chen
Institution
Pennsylvania State University