Major
Chemistry
Anticipated Graduation Year
2022
Access Type
Open Access
Abstract
PhLP-3 is a protein within the thioredoxin-superfamily, and found in eukaryotes of ranging complexity. It has a central thioredoxin domain, containing the redox active site. The thioredoxin domain is framed by an N-terminus triple-helix and a short C-terminus tail. PhLP-3 is hypothesized to function as co-chaperone for the folding of cytoskeletal proteins actin and tubulin. It has been reported that the overexpression or the absence of this protein can lead to abnormal cell morphology and cellular division in various organisms; however, the cellular function of PhLP-3 is still undetermined. Our recent findings on biochemical characterization on malaria PhLP-3 has led to the hypothesis that the protein contains a thioredoxin domain, which harbors the redox active cite to facilitate its function. We expanded our studies to homologous PhLP-3 proteins of human, Drosophila, C. elegans, and Zebrafish. These have been proven through amino acid sequence alignment to have highly conserved PhLP-3. Our experimental results indicate that the redox activity is conserved between these diverse species. Previously, our lab expressed and purified Plasmodium PhLP-3 in high concentrations, but was unsuccessful in crystallization. It was hypothesized that the inability to crystallize resulted from the presence of the 3 helices, leading to obstruction during crystallization and resulting in precipitation. We therefore truncated the 3 helices and worked to crystallize only the thioredoxin domain. We believe that truncating this N-terminus helix domain will improve conditions for crystallization.
Faculty Mentors & Instructors
Dr. Stefan Kanzok, Biology Department
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
PhLP-3 Structure and Function: DmPhLP-3
PhLP-3 is a protein within the thioredoxin-superfamily, and found in eukaryotes of ranging complexity. It has a central thioredoxin domain, containing the redox active site. The thioredoxin domain is framed by an N-terminus triple-helix and a short C-terminus tail. PhLP-3 is hypothesized to function as co-chaperone for the folding of cytoskeletal proteins actin and tubulin. It has been reported that the overexpression or the absence of this protein can lead to abnormal cell morphology and cellular division in various organisms; however, the cellular function of PhLP-3 is still undetermined. Our recent findings on biochemical characterization on malaria PhLP-3 has led to the hypothesis that the protein contains a thioredoxin domain, which harbors the redox active cite to facilitate its function. We expanded our studies to homologous PhLP-3 proteins of human, Drosophila, C. elegans, and Zebrafish. These have been proven through amino acid sequence alignment to have highly conserved PhLP-3. Our experimental results indicate that the redox activity is conserved between these diverse species. Previously, our lab expressed and purified Plasmodium PhLP-3 in high concentrations, but was unsuccessful in crystallization. It was hypothesized that the inability to crystallize resulted from the presence of the 3 helices, leading to obstruction during crystallization and resulting in precipitation. We therefore truncated the 3 helices and worked to crystallize only the thioredoxin domain. We believe that truncating this N-terminus helix domain will improve conditions for crystallization.