Anabaena Sensory Rhodopsin Membrane Protein

Jacquelyn Amadeo-Ranch, Chemistry Undergraduate Student

College of Natural Resources & Sciences

Rhodopsins are light-sensitive proteins within the G-protein coupled receptor (GPCR) family that convert photons into intracellular chemical signals that perform downstream biological functions. Anabaena sensory rhodopsin (ASR) with a 6x-His tag was isolated and purified using Ni-NTA affinity chromatography after evaluating clones of induced E. coli transformed with a pET15b plasmid. The most viable clones discovered from this study were BU3E and Box A. SDS-PAGE gel electrophoresis confirmed the protein size of 20.65 kDa. These results indicate that these 2 strains can be utilized for further studies on ASR function in nanodiscs.

Anabaena Sensory Rhodopsin nanodisc assisted antifungal transport into Brewer's yeast

College of Natural Resources & Sciences

Our experiment hopes to answer the question of whether or not nanodiscs make an effective means of transporting medication, specifically in the treatment of fungal infections. We hope this adds to research that is currently examining its effectiveness in the transport of chemotherapy drugs to prevent collateral cell death; there is current evidence to support these assumptions. To do this, we are testing the transport of Terbinafine into Brewer's yeast in the hopes of inducing cell death.

Anabaena Sensory Rhodopsin Nanodiscs to Probe Transcriptional Regulation

College of Natural Resources & Sciences

The membrane protein Anabaena Sensory Rhodopsin (ASR) is a prokaryotic retinal containing photoactive protein, from Anabaena sp. PCC 7120, undergoes a conformational change upon absorption of light. This causes the release of an associated so-called transducer protein ASRT. It has been proposed that this protein complex directly controls transcription of the cpc genes. The goal of this project is to study photo-induced transcriptional regulation properties of ASR and ASRT. To do this we will assemble ASR nanolipoprotein particles or ASR-NLPs. NLPs are unique in that they allow for a membrane protein to be solubilized while still allowing both ends of the membrane protein to be accessed.

Analysis of a G-Protein Coupled Receptor, CB2

David Lopez, Chemistry Undergraduate Student

College of Natural Resources & Sciences

The CB2 G-protein coupled receptors (GPCR) is found in the brainstem & hippocampus and is devoid of psychotropic effects but is less studied than the CB1 receptor. CB2 is inducible in CNS microglia following inflammation or injury, indicating a role in pain response. Here we sought to analyze CB2 using ChimeraX structures and overcome GPCR protein insolubility in extraction. The pET28a-CNR2, plasmid created and transformed into E.coli pLysS, was confirmed by restriction digest. Purification of CB2 micelles was achieved by affinity chromatography with detergent (43 kD). We aim to utilize nanodiscs to stabilize CB2, allowing studies of the molecular underpinnings informing treatment options.

ANALYSIS OF ACIDITY, CO2 AND OXYGEN CONCENTRATION DURING SUMMER UPWELLING CONDITIONS IN HUMBOLDT BAY, NORTHERN CALIFORNIA

Lindsey Fischer, Oceanography Undergraduate Student

College of Natural Resources & Sciences

This project took place over summer of 2023. We set sensors in Humboldt bay in June and again in August leaving them out for three days. These sensors tracked alkalinity, temperature, and oxygen levels within the water. From this we could look at how summer upwelling changed the water chemistry from June to August.

Analysis of Herbicides on Culturally Significant Plants Throughout Yurok Ancestral Territory

College of Natural Resources & Sciences

Use of herbicides by public and private entities throughout the Yurok ancestral territory, has raised concern for the health of Native peoples exposed to these organic residues on plant materials due to the plants significant role within the Yurok culture ( such as baskets, ceremonial, and medical purposes). Using organic extraction followed by High Performance Liquid Chromatography (HPLC) analysis, the herbicides 2,4-D and Triclopyr were identified on plant material collected from the Ah-Pah location. Continuation of this research involves computational research on 2,4-D derivatives and more sampling sites added for herbicide analysis throughout the Yurok territory.

Analysis of Herbicides on Culturally Significant Plants Throughout Yurok Ancestral Territory

College of Natural Resources & Sciences

Use of herbicides by public and private entities throughout the Yurok ancestral territory, has raised concern for the health of Native peoples exposed to these organic residues on plant materials due to the significant roles that plants have within the Yurok culture (such as basket, medicinal and ceremonial purposes). The herbicides 2,4-D and Triclopyr are analyzed by using organic extraction followed by High Performance Liquid Chromatography (HPLC) analysis. Additional research was also conducted such as computational calculations on 2,4-D and its derivatives along with more sampling sites added to this herbicide research.

Analysis of Post Mortem Human Muscle Proteome via Gel Electrophoresis

College of Natural Resources & Sciences

The changes in proteome human muscle tissue were analyzed using protein extraction and quantification techniques, SDS-PAGE, and two-dimensional gel electrophoresis. Muscle tissue samples were collected at defined postmortem intervals from a single body at the Forensic Investigation Research Station (FIRS) in Grand Junction, CO. These techniques were used to identify decomposition products of the rectus femoris muscle proteins that occur ​post mortem. The ultimate goal of this research is to correlate protein decomposition product masses (via mass spectrometry analysis) to distinctive postmortem intervals measured in accumulated degree days (ADD).

Analysis of Protein Nanodisc Assembly Methods

College of Natural Resources & Sciences

Nanodics which are nanolipoprotein particles (NLPs) can be assembled in order to solubilize and study membrane proteins in a water soluble discoidal particle.. Here, NLPs were assembled using two techniques: dialysis & biobead adsorption. These methods differ in the removal of a detergent called cholate. The Biobead method is much faster, but is relatively untested, whereas the dialysis method is in current use, but is fairly time consuming. By demonstrating the effectiveness of both techniques for nanodisc assembly, solubilization of membrane proteins can be optimized. This will ultimately be helpful in studying ASR, a retinal containing membrane protein for Anabaena (Now Nostoc) PCC 7120.

Analysis of Trace Metals in Seawater using Pre-Concentration Techniques and Flame Atomic Absorption Spectroscopy

College of Natural Resources & Sciences

Many trace metals are essential micronutrients for phytoplankton, and despite their low concentrations in seawater, trace metals can have large impacts on biological processes. Also due to their low concentrations, the measurement of trace metal concentrations requires precise analytical techniques. One common method utilizes expensive instrumentation that we do not have at HSU. In this research project, we adapted published methods to use the Flame Atomic Absorption Spectrometer, an instrument in the HSU Chemistry Department. Results show successful adaptation of the method for zinc and manganese. Current work streamlines the method to allow for 8-fold faster sample preparation.