Project Title
Unveiling the crystal structure of lipid-bound HALT-1: Investigating pore formation mechanisms for enhanced control of inflammation and therapeutic development
Research Project
Unveiling the crystal structure of lipid-bound HALT-1: Investigating pore formation mechanisms for enhanced control of inflammation and therapeutic development
Project Description
We have previously isolated Hydra actinoporin-like toxin 1 (HALT-1) from Hydra magnipapillata, a freshwater cnidarian species. HALT-1 exhibits preferential binding to sulfatide-associated membranes, leading to the formation of pores on human cells. This pore formation results in either mechanical cell disruption or programme cell death, depending on the dosage of HALT-1 applied to human cells. We have conjugated HALT-1 with a single chain variable fragment (scFv) antibody, producing a recombinant conjugate in an E. coli system. This conjugate demonstrated effective reduction in the viability of pro-inflammatory M1 macrophages, highlighting its potential in controlling inflammation in various inflammatory diseases. However, the precise mechanism underlying HALT-1's pore formation remains poorly understood. In this proposed study, our objective is to elucidate the key residues in HALT-1 responsible for lipid membrane recognition, oligomerization, and membrane insertion. To achieve this, we will create a HALT-1-liposome complex and employ X-ray crystallography to examine detailed interactions between HALT-1 and the liposome. By performing alanine substitution and assessing cytotoxicity loss, we will confirm the crucial role of these identified residues. Our research aims to gain molecular insights into pore formation, thereby enhancing the specificity and efficacy of HALT-1-scFv. This will be achieved by removing HALT-1's lipid-binding ability in scFv-HALT-1, directing binding solely through scFv and increasing specificity. Additionally, through modification of relevant residues in HALT-1 to favor pore formation, we can enhance the cytotoxicity of scFv-HALT-1.
Vacancy For
Graduate Research Assistant
Number of Vacancies
1
Duration
2 years
Requirements
1. Completed a Bachelor degree in the relevant and related field with a minimum CGPA of 3.5
2. Keen interest in proteomics, particularly in protein purification and crystallization techniques
3. Ability to collaborate with researchers from other universities
4. Full-time enrollment into Master's programme in School of Medical and Life Sciences, Sunway University. (Tuition Fee Waiver provided)
To apply, please write to:
Dr Hwang Jung Shan
Email: [email protected]
