Relative Spatial Position of a Snake Neurotoxin and the Reduced Disulfide Bond α(Cys192-Cys193) at the αγ Interface of the Nicotinic Acetylcholine Receptor
We determined the distances separating five functionally important residues (Gln 10 , Lys 27 , Trp 29 , Arg 33 , and Lys 47 ) of a three-fingered snake neurotoxin from the reduced disulfide bond α(Cys 192 -Cys 193 ) located at the αγ interface of the Torpedo nicotinic acetylcholine receptor. Each...
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Published in | The Journal of biological chemistry Vol. 275; no. 33; p. 25608 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
American Society for Biochemistry and Molecular Biology
18.08.2000
|
Online Access | Get full text |
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Summary: | We determined the distances separating five functionally important residues (Gln 10 , Lys 27 , Trp 29 , Arg 33 , and Lys 47 ) of a three-fingered snake neurotoxin from the reduced disulfide bond α(Cys 192 -Cys 193 ) located at the αγ interface of the Torpedo nicotinic acetylcholine receptor. Each toxin position was substituted individually for a cysteine, which was then linked
to a maleimido moiety through three different spacers, varying in length from 10 to 22 Ã
. We estimated the coupling efficiency
between the 15 toxin derivatives and the reduced cystine α(192â193) by gel densitometry of Coomassie Blue-stained gels. A
nearly quantitative coupling was observed between αCys 192 and/or αCys 193 and all probes introduced at the tip of the first (position 10) and second (position 33) loops of Naja nigricollis α-neurotoxin. These data sufficed to locate the reactive thiolate in a âcroissant-shapedâ volume comprised between the first
two loops of the toxin. The volume was further restrained by taking into account the absence or partial coupling of the other
derivatives. Altogether, the data suggest that αCys 192 and/or αCys 193 , at the αγ interface of a muscular-type acetylcholine receptor, is (are) located in a volume located between 11.5 and 15.5
Ã
from the α-carbons at positions 10 and 33 of the toxin, under the tip of the toxin first loop and close to the second one. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M002362200 |