Oceanic life is facing the deleterious aftermath of coral bleaching. To reverse the damages introduced by anthropological means, it is imperative to study fundamental properties of corals. One way to do so is to understand the metabolic pathways and protein functions of corals that contribute to the resilience of coral reefs. Although genomic sequencing and structural modeling has yielded significant insights for well-studied organisms, more investigation must be conducted for corals. Better yet, quantifiable experiments are far more crucial to the understanding of corals. The objective is to clone, purify, and assess coral proteins from the cauliflower coral species known as Pocillopora damicornis. Presented here is the pipeline for how 3-D structural modeling can help support the experimental data from studying soluble proteins in corals. Using a multi-step selection approach, 25 coral genes were selected and retrieved from the genomic database. Using Escherischia coli and Homo sapiens homologues for sequence alignment, functional properties of each protein were predicted to aid in the production of structural models. Using D-SCRIPT, potential pairwise protein-protein interactions (PPI) were predicted amongst these 25 proteins, and further studied for identifying putative interfaces using the ClusPro server. 10 binding pockets were inferred for each pair of proteins. Standard cloning strategies were applied to express 4 coral proteins for purification and functional assays. 2 of the 4 proteins had visible bands on the Coomassie stained gel and were able to advance to the purification step. Both proteins exhibited a faint band at the expected migration distance for at least one of the elutions. Finally, PPI was carried out by mixing protein samples and running in a native gel, resulting in one potential pair of PPI.
Diisobutylene maleic acid, or DIBMA, offers a novel approach to integral membrane protein extraction without requiring the use of detergent. This copolymer extracts the protein along with the surrounding lipids, creating native nanodiscs. This method of solubilization is the preferred method, as traditional detergent solubilization can possibly alter the structural and functional integrity of the membrane protein. DIBMA solubilization, on the other hand, is able to create a more stable environment for the integral membrane protein, while allowing purification through commonly used chromatography methods similar to established detergent solubilization protocols. In this project, we study the ability of DIBMA to isolate the integral membrane protein, chloroplast ATP synthase, without the use of detergents.
Purification, Characterization, and Structural Determination of Proteins Vital to Infectious Disease
Aging is a universal process that every being encounters on their journey of life. The effect of dance as a form of improvement of physical and well-being on the aging body brings upon the question of the impact of somatic-based movement, specifically gestural movement on the perceptions of aging within older and younger adults through a bi-cultural lens of the United States and India. This was investigated using a series of creative partnership workshops that included listening, drawing, and culminating movement activities, followed by a group discussion about the creative process. There were four different participant groups: a group of college students taking an Aging in American Culture collegiate class, a Somatic Practices collegiate dance class, a group of older adults at the Tempe Multigenerational Center, and a group of older adults in Bangalore, India. Inter-generational and cross-cultural observations were discussed, and it was seen that the workshops were able to foster feelings of community and camaraderie among participants depending on the group’s relationship to dance-making. From the research, a dance performance was developed and performed specifically with women of color in ASU’s Dance program with the choreographic process discussed in detail, along with key takeaways about facilitating a multi-dimensional experience for the dancers and choreographer. Future directions for the work include working with intergenerational populations and researching community effects on gendered aging experiences across cultures in the dance world.