NMR에 의한 anti-Ex-A IgG의 항원결합부위 해석

• 영문명: Paratope Mapping of Anti-Ex-A IgG as Studied by NMR
• 발행기관: 대한약학회
• 저자명: 김하형(Ha Hyung Kim) 이광표(Kwang Pyo Lee) 가토 코이치(Koichi Kato) 아라타 요우지(Yoji Arata)
• 간행물 정보: 『약학회지』제40권 제4호 (1996년), 422~427쪽, 전체 6쪽
• 주제분류: 의약학 > 기타의약학
• 파일형태: PDF
• 발행일자: 1996.08.31

영문 초록

The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-d2, L-Phe-d5, L-Trp-d5, L-Tyr-2,6-d2 and L-[1-13C]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using 13C-15N double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the VH domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.

키워드

anti-Ex-A IgG paratope mapping NMR stable-isotope photo CIDNP.