| 학회 | 한국고분자학회 |
| 학술대회 | 2005년 가을 (10/13 ~ 10/14, 제주 ICC) |
| 권호 | 30권 2호 |
| 발표분야 | 의료용 고분자 부문위원회 |
| 제목 | Direct measurement of molecular interaction between Lipopolysaccharide and CD14,LBP |
| 초록 | We used atomic force microscopy (AFM) to explore the binding forces of immune protein(LBP, CD14) and Lipopolysaccharide(LPS). To detect single molecular recognition events(1) , genetically engineered histidine-tagged immune protein was coupled onto AFM tips modified with self-assembled monolayers (SAMs) of nitrilotriacetic acid- and hydroxyl group terminated alkanethiols while LPS was supported by lipid bilayer on mica. The quality of the protein functionalization procedure on AFM tip was validated using X-ray photoelectron spectroscopy (surface chemical composition), FT-IR (chemical bonding of linker), surface plasmon resonance (SPR, specific binding in aqueous solution and binding affinity) (2) , Matrix Assisted Laser Desorption Ionization Time of flight (MALDI-TOF, protein immobilization) and Field emission scanning electron microscope (FE-SEM, Tip radius of curvature). LPS was immobilized to the model biomembrane including lipid bilayer for more similar natural membrane, and model biomembrane has been prepared by Langmiur - Blodgett(LB) trough. With these probe and substrate, we have measured the adhesion force between CD14, LBP and LPS by changing the loading rate using AFM. Kinetic parameters obtained from the relation between the adhesion force and loading rate could be used to thermodynamically characterize the binding behavior of these molecules. Additionally, as many reports show, the directly measured force will be used to explore the energy landscape of ligand-receptor unbinding. (3)(4) Reference (1)S.Allen, S.M. Rigby-Singleton, and P.O'Shea, Biochemical Society Transaction , 2003, 31, 1052 (2)Alexandre Berquand, Nan Xia, and Yves F.Dufrene , Langmuir, 2005,21,5517 (3)Bell, G.I., Science. 1978, 200, 618. (4)Evans,E., and Ritchie,K., Biophys.J. ,1997, 72, 1541 |
| 저자 | 김종수1, 장순남2, 김의숙2, 김성수2, 조길원2 |
| 소속 | 1포항공과대, 2포항공대 |
| 키워드 | AFM; Dynamic Force Spectroscopy; LPS; LBP; CD14 |
