Life Sciences & Biotechnology
Title : | Understanding on/off kinetics of LOV-domain proteins to construct optogenetic tools |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Rajiv Kumar Kar, Indian Institute Of Technology (IIT) Guwahati, Assam |
Timeline Start Year : | 2022 |
Timeline End Year : | 2024 |
Contact info : | rajivkar@iitg.ac.in |
Equipments : | Data processing workstation
Water Bath
Laminar air flow hood
Double door refrigerator
GPU Graphics card
Magnetic stirrer bath |
Details
Executive Summary : | The light-activation of biological macromolecules enables researchers to develop optogenetic tools for monitoring and controlling cellular events. The global optogenetic market is expected to grow significantly, with international projects expected to reach $59 billion by 2026. This proposal aims to harness the fundamental mechanism of photoreceptor proteins to repurpose biological phenomena. The LOV-domain is considered a non-neural optogenetic candidate that can control macromolecular location with spatiotemporal resolution. The project aims to answer the fundamental question of how amino-acid networks manipulate light-activation/deactivation kinetics in LOV proteins. The primary step in this process is to understand comprehensive photochemistry upon blue-light irradiation in LOV-containing proteins. Experiments will involve absorbance spectra, steady-state fluorescence measurement, redox titration, and hybrid quantum mechanics/molecular mechanics QM/MM dynamics for ground and excited state electronic structures. The study will involve three proteins with variable kinetics: Avena sativa phototropin-1 AsLOV2, Bacillus subtilis stress response protein YtvA, and fungal circadian clock photoreceptor from Neurospora crassa VVD. The project's central goal is to understand the photocycle kinetics of the LOV domain to design protein mutants with desired spectroscopic properties. The project will explore the relationship of atomistic insights with optical properties to correlate with photocycle kinetics. The proposal aims to design LOV mutants for developing technologies such as voltage-sensitive fluorescent proteins and light techniques like cardioversion, protein tracking, and resolution. |
Total Budget (INR): | 28,08,340 |
Organizations involved