Novel unconventional binding site in the variable region of immunoglobulins

被引:40
作者
Rajagopalan, K
Pavlinkova, G
Levy, S
Pokkuluri, PR
Schiffer, M
Haley, BE
Kohler, H
机构
[1] UNIV KENTUCKY, DEPT MICROBIOL & IMMUNOL, LEXINGTON, KY 40536 USA
[2] UNIV KENTUCKY, COLL PHARM, DIV MED CHEM & PHARMACEUT, LEXINGTON, KY 40536 USA
[3] UNIV KENTUCKY, MARKEY CANC CTR, LEXINGTON, KY 40536 USA
[4] STANFORD UNIV, SCH MED, DIV ONCOL, STANFORD, CA 94305 USA
[5] ARGONNE NATL LAB, CTR MECH BIOL, ARGONNE, IL 60439 USA
关键词
antibody; nucleotide binding site; affinity linker; computer model;
D O I
10.1073/pnas.93.12.6019
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The variable immunoglobulin (Ig) domains contain hypervariable regions that are involved in the formation of the antigen binding site. Besides the canonical antigen binding site, so-called unconventional sites also reside in the variable region that bind bacterial and viral proteins, Docking to these unconventional sites does not typically interfere with antigen binding, which suggests that these sites may be a part of the biological functions of Tgs. Herein, a novel unconventional binding site is described, The site is detected with 8-azidopurine nucleotide photoaffinity probes that label antibodies efficiently and under mild conditions. Tryptic peptides were isolated from photolabeled monoclonal antibodies and aligned with the variable antibody domains of heavy and light chains, The structure of a variable Ig fragment was used to model the binding of the purine nucleotide to invariant residues in a hydrophobic pocket of the Ig molecule at a location distant from the antigen binding site. Monoclonal and polyclonal antibodies were biotinylated with the photoaffinity linker and used in fluorescence-activated cell sorter and ELISA analyses. The data support the utility of this site for tethering diagnostic and therapeutic agents to the variable Ig fragment region without impairing the structural and functional integrity of antibodies.
引用
收藏
页码:6019 / 6024
页数:6
相关论文
共 27 条
[1]  
ANDRIA ML, 1990, J IMMUNOL, V144, P2614
[2]   IMMUNOGLOBULIN-V(H)3 GENE-PRODUCTS - NATURAL LIGANDS FOR HIV GP120 [J].
BERBERIAN, L ;
GOODGLICK, L ;
KIPPS, TJ ;
BRAUN, J .
SCIENCE, 1993, 261 (5128) :1588-1591
[3]   PROTEIN DATA BANK - COMPUTER-BASED ARCHIVAL FILE FOR MACROMOLECULAR STRUCTURES [J].
BERNSTEIN, FC ;
KOETZLE, TF ;
WILLIAMS, GJB ;
MEYER, EF ;
BRICE, MD ;
RODGERS, JR ;
KENNARD, O ;
SHIMANOUCHI, T ;
TASUMI, M .
JOURNAL OF MOLECULAR BIOLOGY, 1977, 112 (03) :535-542
[4]  
Brunger A. T., 1992, X PLOR SYSTEM X RAY
[5]  
Cleary Patrick, 1994, Trends in Microbiology, V2, P131, DOI 10.1016/0966-842X(94)90600-9
[6]   SETOR - HARDWARE-LIGHTED 3-DIMENSIONAL SOLID MODEL REPRESENTATIONS OF MACROMOLECULES [J].
EVANS, SV .
JOURNAL OF MOLECULAR GRAPHICS, 1993, 11 (02) :134-&
[7]   MOLECULAR-BASIS OF ANTIGEN MIMICRY BY AN ANTI-IDIOTOPE [J].
FIELDS, BA ;
GOLDBAUM, FA ;
YSERN, X ;
POLJAK, RJ ;
MARIUZZA, RA .
NATURE, 1995, 374 (6524) :739-742
[8]   IMMUNOGLOBULIN HEAVY-CHAIN BINDING-PROTEIN [J].
HAAS, IG ;
WABL, M .
NATURE, 1983, 306 (5941) :387-389
[9]  
HALEY BE, 1991, METHOD ENZYMOL, V200, P477
[10]  
HARLOW E, 1988, ANTIBODIES LABORATOR, P423