Screening of Lipoprotein Lipase Regulators based on ApolipoproteinC2 Binding Site

• Main Authors: Yu-Zong Guan, 管昱宗
• Other Authors: Nai-Wan Hsiao
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/25470416316777298666

碩士 === 國立彰化師範大學 === 生物技術研究所 === 102 === Lipoprotein lipase (LPL) is a key enzyme in the lipid metabolism. When chylomicrons are generated in the small intestine, they carry large amounts of triglycerides (TGs) decomposed from food. TGs were transported to the bloodstream and hydrolyzed by LPL into fatty acids which are used in various tissues. Moreover, a TG in liver will be assembled with lipoproteins and ApoC-II to form a multi-molecule lipoprotein which is exported to the blood and then hydrolyze via LPL. The lipid metabolism strongly correlates with the energy used in tissues, so the hydrolyzed function of LPL is important in the energy application in the body. Irregular lipid metabolism will lead atherosclerosis, obesity, hyperlipidemia, etc. Thus, appropriate regulation of LPL may prevent these diseases. LPL has hydrolyzed function only in the homodimer state, and the binding of heparin sulphate-proteoglycans (HSPG) and apolipoprotein (ApoC-II) will assist LPL in hydrolysis. Our design is focused on the sites that may affect the activity of the protein. We have conducted the activity tests of small molecules, peptides, and antibodies. The synthetic tetrapeptide KGEE was demonstrated having inhibition activity to LPL, and also confirmed ApoC-II all-length in LPL enzyme assay has the effect of active. In the antibody experiments, 2F5 (HIV-1 Neutralizing antibody) was found by using the computer simulation. The screening of peptides and antibodies operating and simulate the human lipoprotein lipase structure through computer aid program. Identify suitable antibodies from antibody libraries based the structure and sequence alignment of ApoC-II binding site with antigen 。After biopannimg of peptides and antibodies library followed ELISA test to identify the sequence affinity well。Analysis and observation the interaction with peptides where binding site by computer aid software .The experimental process from design to final analyzes with computer-assisted make the experimental data more effectively.