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  • 09월 04일 17시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제114회 대한화학회 학술발표회, 총회 및 기기전시회 안내 NMR Structural Analysis of Antimicrobial peptides, LPcin analogs with Enhanced Activities

2014년 8월 28일 17시 24분 47초
ANAL.P-579 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
10월 15일 (수요일) 16:00~19:00
저자 및
김경섭, 정지호, 김지선, 김용애*
한국외국어대학교 화학과, Korea
Lactophoricin (LPcin), a cationic amphipathic peptide consists of 23-mer peptide, corresponds to the carboxy terminal 113-135 region of Component-3 of bovine proteose-peptone PP3. LPcin was designed and modified to enhance antibacterial activity using mutation, sequence shuffling for better amphipathic characteristics. It helps to improve interacting with bacterial membranes. Three candidates were selected among 12 LPcin analogs which have better antibacterial activity than LPcin. We named them LPcin-yk1, LPcin-yk2 and LPcin-yk3 respectively. LPcin-yk1 and LPcin-yk3 has shorter amino acids at C-terminus, LPcin-yk2 has same number of amino acids but several amino acids were substituted for better amphipathic characteristics. To understand the correlation between their structures and antibacterial mechanism, we studied how the modified conformation of LPcin analogs can affect their antimicrobial activity using various biophysical techniques. We successfully overexpressed in the form of fusion protein in Escherichia coli and purified with biophysical techniques such as Ni-NTA affinity chromatography, dialysis, CNBr chemical cleavage and reversed-phase semi-prep.HPLC. To verify the purified peptides, we performed Tris-tricine PAGE, MALDI-TOF MS and CD spectrometry. In order to investigate the structures and dynamics, we use various 1D and 2D NMR techniques using micelle and bicelle samples to make membrane environments. In here, we will present not only the optimization of high-yield expression and purification, but also solution and solid-state NMR experiments to elucidate their structures. The structural calculations of LPcin analogs using Discovery Studio 3.1 and tilt angle prediction based on SAMPI4 Solid state NMR spectra and PISA Wheel pattern were also underway.