How protein recognizes ladder-like polycyclic ethers - Interactions between ciguatoxin (CTX3C) fragments and its specific antibody 10C9

被引:15
作者
Ui, Mihoko [1 ]
Tanaka, Yoshikazu [1 ]
Tsumuraya, Takeshi [3 ]
Fujii, Ikuo [3 ]
Inoue, Masayuki [2 ,4 ]
Hirama, Masahiro [4 ]
Tsumoto, Kouhei [1 ]
机构
[1] Univ Tokyo, Grad Sch Frontier Sci, Dept Med Genome Sci, Chiba 2778562, Japan
[2] Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 113003, Japan
[3] Osaka Prefecture Univ, Grad Sch Sci, Dept Biol Sci, Osaka 5998570, Japan
[4] Tohoku Univ, Grad Sch Sci, Dept Chem, Sendai, Miyagi 9808578, Japan
关键词
D O I
10.1074/jbc.M801282200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Ciguatoxins are a family of marine toxins composed of transfused polycyclic ethers. It has not yet been clarified at the atomic level on the pathogenic mechanism of these toxins or the interaction between a polycyclic ether compounds and a protein. Using the crystal structures of anti-ciguatoxin antibody 10C9 Fab in ligand-free form and in complexes with ABCD-ring (CTX3C-ABCD) and ABCDE-ring (CTX3C-ABCDE) fragments of the antigen CTX3C at resolutions of 2.6, 2.4, and 2.3 angstrom, respectively, we elucidated the mechanism of the interaction between the polycyclic ethers and the antibody. 10C9 Fab has an extraordinarily large and deep binding pocket at the center of the variable region, where CTX3C-ABCD or CTX3C-ABCDE binds longitudinally in the pocket via hydrogen bonds and van der Waals interactions. Upon antigen-antibody complexation, 10C9 Fab adjusts to the antigen fragments by means of rotational motion in the variable region. In addition, the antigen fragment lacking the E-ring induces a large motion in the constant region. Consequently, the thermostability of 10C9 Fab is enhanced by 10 degrees C upon complexation with CTX3C-ABCDE but not with CTX3C-ABCD. The crystal structures presented in this study also show that 10C9 Fab recoginition of CTX3C antigens requires molecular rearrangements over the entire antibody structure. These results further expand the fundamental understanding of the mechanism by which ladder-like polycyclic ethers are recognized and may be useful for the design of novel therapeutic agents by antibodies, marine toxins, or new diagnostic reagents for the detection and targeting of members of the polycyclic ether family.
引用
收藏
页码:19440 / 19447
页数:8
相关论文
共 38 条
[1]  
[Anonymous], ACTA CRYSTALLOGR
[2]   MOLECULAR-BASIS OF CROSS-REACTIVITY AND THE LIMITS OF ANTIBODY-ANTIGEN COMPLEMENTARITY [J].
AREVALO, JH ;
TAUSSIG, MJ ;
WILSON, IA .
NATURE, 1993, 365 (6449) :859-863
[3]   Anatomy of an antibody molecule: structure, kinetics, thermodynamics and mutational studies of the antilysozyme antibody D1.3 [J].
Braden, BC ;
Goldman, ER ;
Mariuzza, RA ;
Poljak, RJ .
IMMUNOLOGICAL REVIEWS, 1998, 163 :45-57
[4]   Crystallography & NMR system:: A new software suite for macromolecular structure determination [J].
Brunger, AT ;
Adams, PD ;
Clore, GM ;
DeLano, WL ;
Gros, P ;
Grosse-Kunstleve, RW ;
Jiang, JS ;
Kuszewski, J ;
Nilges, M ;
Pannu, NS ;
Read, RJ ;
Rice, LM ;
Simonson, T ;
Warren, GL .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1998, 54 :905-921
[5]  
DAVIES DR, 1990, ANNU REV BIOCHEM, V59, P439, DOI 10.1146/annurev.biochem.59.1.439
[6]   Engineering stability into Escherichia coli secreted Fabs leads to increased functional expression [J].
Demarest, Stephen J. ;
Chen, Gang ;
Kimmel, Bruce E. ;
Gustafson, David ;
Wu, Jane ;
Salbato, Jared ;
Poland, John ;
Elia, Marikka ;
Tan, Xuqiu ;
Wong, Ken ;
Short, Jay ;
Hansen, Genevieve .
PROTEIN ENGINEERING DESIGN & SELECTION, 2006, 19 (07) :325-336
[7]   THE PRICE OF LOST FREEDOM - ENTROPY OF BIMOLECULAR COMPLEX-FORMATION [J].
FINKELSTEIN, AV ;
JANIN, J .
PROTEIN ENGINEERING, 1989, 3 (01) :1-3
[8]   A broad range of Fab stabilities within a host of therapeutic IgGs [J].
Garber, Ellen ;
Demarest, Stephen J. .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2007, 355 (03) :751-757
[9]   Humanization of the mouse anti-Fas antibody HFE7A and crystal structure of the humanized HFE7A Fab fragment [J].
Haruyama, H ;
Ito, S ;
Miyadai, K ;
Takahashi, T ;
Kawaida, R ;
Takayama, T ;
Hanzawa, H ;
Hata, T ;
Yamaguchi, J ;
Yoshida-Kato, H ;
Ichikawa, K ;
Ohsumi, J ;
Yonehara, S ;
Serizawa, N .
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 2002, 25 (12) :1537-1545
[10]   Total synthesis of ciguatoxin CTX3C: A venture into the problems of ciguatera seafood poisoning [J].
Hirama, M .
CHEMICAL RECORD, 2005, 5 (04) :240-250