Date of Award
2025
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Chemistry
Committee Chair
Ifedayo Victor Ogungbe
Committee Member
Pam Twigg
Research Advisor
Marc Pusey
Subject(s)
Neovascularization inhibitors, Proteins--Purification, Gene expression, Angiopoietin-like protein 4 (ANGPTL4)
Abstract
This research focused on developing protocols to express and purify the N-terminus of angiopoietin-like protein 4 (nANGPTL4), and identify its structural characteristics to understand its possible contribution to cancer metastasis. Bacteria expression was optimized by adding magnesium, calcium, and glycerol to the initial Luria-Bertani (LB) broth incubation, then transferring to Terrific Broth (TB) for induction. Purification was enhanced through overnight nickel resin suspension, extended sonification, and buffer adjustments. Protein recovery was further improved by transferring size exclusion columns to G50 superfine resin. Protein purity was analyzed using SDS-PAGE and native gel electrophoresis, while protein content was determined through UV-Vis spectroscopy. Circular dichroism and 1D 1H NMR data suggested the presence of alpha helices in nANGPTL4. The research provides a foundation for future studies, with ongoing optimization of protein crystal screens for X-ray diffraction or microED analysis. These findings contribute to understanding nANGPTL4’s potential role in tumor progression and metastasis inhibition.
Recommended Citation
Paskach, Alexis Leann, "Expression, purification, and characterization of the angiopoietin-like 4 protein N-terminus" (2025). Theses. 774.
https://louis.uah.edu/uah-theses/774