Structural Biology Commons^™

Computational Investigations Into Binding Dynamics Of Tau Protein Antibodies: Using Machine Learning And Biophysical Models To Build A Better Reality, Katherine Lee

University Scholar Projects

Misregulation of post-translational modifications of microtubule-associated protein tau is implicated in several neurodegenerative diseases including Alzheimer’s disease. Hyperphosphorylation of tau promotes aggregation of tau monomers into filaments which are common in tau-associated pathologies. Therefore, tau is a promising target for therapeutics and diagnostics. Recently, high-affinity, high-specificity single-chain variable fragment (scFv) antibodies against pThr-231 tau were generated and the most promising variant (scFv 3.24) displayed 20-fold increased binding affinity to pThr-231 tau compared to the wild-type. The scFv 3.24 variant contained five point mutations, and intriguingly none were in the tau binding site. The increased affinity was hypothesized to occur due …

Development Of A Sonically Powered Biodegradable Nanogenerator For Bone Regeneration, Avi S. Patel

University Scholar Projects

Background: Reconstruction of bone fractures and defects remains a big challenge in orthopedic surgery. While regenerative engineering has advanced the field greatly using a combination of biomaterial scaffolds and stem cells, one matter of difficulty is inducing osteogenesis in these cells. Recent works have shown electricity’s ability to promote osteogenesis in stem cell lines when seeded in bone scaffolds; however, typical electrical stimulators are either (a) externally housed and require overcomplex percutaneous wires be connected to the implanted scaffold or (b) implanted non-degradable devices which contain toxic batteries and require invasive removal surgeries.

Objective: Here, we establish a biodegradable, piezoelectric …

Computational Investigations Into The Molecular Underpinnings Of Eyesight Signaling Pathways, Shaan Kamal

University Scholar Projects

Phosphodiesterase 6 (PDE6) is a critical enzyme in the eyesight-signaling pathway. When activated, PDE6 hydrolyzes cGMP to GMP, which deactivates cGMP- gated ion channels, causing hyperpolarization of the cell and activating the sensory neurons responsible for vision. Within the PDE family, PDE6 is the only enzyme known to have an inhibitory subunit (PDE6-γ), which allows for the regulation of cGMP levels. When PDE6-γ is bound to PDE6, the enzyme is turned “off” and cannot catalyze cGMP. The α subunit of the G-protein transducin removes PDE6-γ and activates PDE6. PDE6 has proven problematic to isolate, making it difficult to study experimentally …

Optimization Of Expression And Purification Of The Rig-I-Like-Receptor, Lgp2, Srinath-Reddi Pingle

University Scholar Projects

The innate immune system is one of the first lines of defense against pathogens. RIG-I like receptors (RLRs) are a class of cytosolic receptors that recognize molecular patterns of invading pathogens and signal for downstream interferon induction. The RLR family consists of three proteins, RIG-I, MDA5, and LGP2. MDA5 and LGP2 work together to recognize and bind long viral dsRNA. LGP2 is thought to regulate MDA5 activation, but little is understood about this process. Characterization of the mechanism of action of these receptors requires careful biophysical and biochemical analyses. I have developed expression and purification of methods LGP2 and MDA5 …

Stability Of Norwalk Virus Capsid Protein Interfaces Evaluated By In Silico Nanoindentation, Prakhar Bansal

University Scholar Projects

Studying the mechanical properties of viral capsids can give several insights into not only the lifecycle of the virus, but also into potential drug targets to thwart the progression of viral infection. Nanoindentation using an atomic force microscope is a useful technique for determining structural properties of small molecules and particles, and is commonly used to study viral capsids. This technique utilizes the probe of the microscope to push down on the capsid and record the forces along the indentation path. We ran this experiment in silico where we simulated the nanoindentation of Norwalk virus capsids using molecular dynamics. Running …

Biophysical Studies Of Styrene-Maleic Acid Copolymer Stabilized Membrane Mimetics, Diane Yu

University Scholar Projects

Traditional methods of isolating membrane proteins involve the use of detergents that destabilize the membrane environment. Biophysical studies of membrane proteins are challenged by the requirement of a stable membrane platform. Lipodisq® nanoparticles are novel membrane mimetics that consist of 3:1 styrene-maleic acid copolymer encircling phospholipids to form discoidal species on the order of 10 nm in diameter. Bacteriorhodopsin was previously incorporated into Lipodisq® nano-particles from its native purple membrane without the addition of detergent. In this study, dynamic light scattering was used to investigate the stability of empty Lipodisq® nano-particles over time. These data show that Lipodisq® nano-particles are …