Life Sciences & Biotechnology
Title : | Unraveling the molecular-to-atomic structure of plant seed storage protein bodies containing amyloidic composites |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Prof. Ashwani Kumar Thakur, Indian Institute Of Technology Kanpur (IITK), Uttar Pradesh |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | akthakur@iitk.ac.in |
Details
Executive Summary : | Plant seed embryos require nutrient availability during germination, and seed storage protein bodies (SSPB) provide this. These bodies contain storage proteins, defense peptides, endoproteases, and minerals. However, the molecular and atomic-level structure of the protein assemblies in SSPB is not yet decoded. A recent discovery revealed that SSPB in monocots and dicots comprises a composite structure of storage proteins in their native globular state and amyloidic assemblies. The packing of individual proteins in their soluble or amyloidic fold in the SSPB remains unexplored. This proposal aims to find the atomic structural details of the individual protein components in their native state and the amyloidic composites of the SSPB using high-resolution techniques like single-particle reconstruction cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET). The intact SSPB and protein composites will be isolated and purified, analyzed for physicochemical characteristics and purity using dynamic light scattering, infra-red spectroscopy, and size exclusion chromatography. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) will be used to ascertain homogeneity and monodispersity of the samples. Cryo-EM will be used to build atomic-level 3D models, while cryo-ET will reconstruct a 3D projection of SSPB inside protoplasts. This proposal will not only elucidate the fundamental molecular-to-atomic-level structure but also shed light on the structure-function relationship of functional amyloids in plant and seed physiological processes. |
Co-PI: | Dr. Appu Kumar Singh, Indian Institute Of Technology Kanpur (IITK), Uttar Pradesh-208016 |
Total Budget (INR): | 40,20,016 |
Organizations involved