Crystal structure of an acetylcholinesterase-fasciculin complex: Interaction of a three-fingered toxin from snake venom with its target

被引:230
作者
Harel, M
Kleywegt, GJ
Ravelli, RBG
Silman, I
Sussman, JL
机构
[1] WEIZMANN INST SCI, DEPT BIOL STRUCT, IL-76100 REHOVOT, ISRAEL
[2] UNIV UPPSALA, CTR BIOMED, DEPT MOLEC BIOL, S-75124 UPPSALA, SWEDEN
[3] BIJVOET CTR BIOMOLEC RES, DEPT CRYSTAL & STRUCT CHEM, 3584 CH UTRECHT, NETHERLANDS
[4] BROOKHAVEN NATL LAB, DEPT BIOL, UPTON, NY 11973 USA
[5] WEIZMANN INST SCI, DEPT NEUROBIOL, IL-76100 REHOVOT, ISRAEL
关键词
alpha/beta hydrolase fold; acetylcholinesterase; fasciculin; protein-protein complex; snake toxins;
D O I
10.1016/S0969-2126(01)00273-8
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
Background: Fasciculin (FAS), a 61-residue polypeptide purified from mamba venom, is a three-fingered toxin which is a powerful reversible inhibitor of acetylcholinesterase (AChE). Solution of the three-dimensional structure of the AChE/FAS complex would provide the first structure of a three-fingered toxin complexed with its target. Results: The structure of a complex between Torpedo californica AChE and fasciculin-II (FAS-II), from the venom of the green mamba (Dendroaspis angusticeps) was solved by molecular replacement techniques, and refined at 3.0 Angstrom resolution to an R-factor of 0.231. The structure reveals a stoichiometric complex with one FAS molecule bound to each AChE subunit. The AChE and FAS conformations in the complex are very similar to those in their isolated structures. FAS is bound at the 'peripheral' anionic site of AChE, sealing the narrow gorge leading to the active site, with the dipole moments of the two molecules roughly aligned. The high affinity of FAS for AChE is due to a remarkable surface complementarity, involving a large contact area (similar to 2000 Angstrom(2)) and many residues either unique to FAS or rare in other three-fingered toxins. The first loop, or finger, of FAS reaches down the outer surface of the thin aspect of the gorge. The second loop inserts into the gorge, with an unusual stacking interaction between Met33 in FAS and Trp279 in AChE. The third loop points away from the gorge, but the C-terminal residue makes contact with the enzyme. Conclusions: Two conserved aromatic residues in the AChE peripheral anionic site make important contacts with FAS. The absence of these residues from chicken and insect AChEs and from butyrylcholinesterase explains the very large reduction in the affinity of these enzymes for FAS. Several basic residues in FAS make important contacts with AChE. The complementarity between FAS and AChE is unusual, inasmuch as it involves a number of charged residues, but lacks any intermolecular salt linkages.
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页码:1355 / 1366
页数:12
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