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Insect nervous system [(3)H]saxitoxin binding sites: characterization and target size.

S C Lummis¹, D B Sattelle

¹AFRC Unit of Insect Neurophysiology and Pharmacology, Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, England.

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|May 28, 2010

Summary

Saxitoxin (STX) binding in cockroach nervous system membranes reveals specific sites on voltage-operated sodium channels. Radiation inactivation estimates the insect sodium channel

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Area of Science:

Neuroscience
Biochemistry
Pharmacology

Background:

Saxitoxin (STX) is a critical tool for studying voltage-operated sodium channels in various organisms.
Cockroach (Periplaneta americana) nervous system membranes exhibit specific [(3)H]STX binding, indicating the presence of these channels.

Purpose of the Study:

To characterize the specific saxitoxin binding sites in the cockroach nervous system.
To determine the molecular size of the insect sodium channel using radiation inactivation.

Main Methods:

Radioligand binding assays using [(3)H]STX.
Scatchard analysis to determine binding affinity (K(D)) and site density (B(max)).
Radiation inactivation to estimate the molecular weight (M(r)) of the target protein.

Main Results:

Specific [(3)H]STX binding was identified with a K(D) of 0.84 nM and B(max) of 8.25 pmol/mg protein.
Binding was inhibited by STX and tetrodotoxin (TTX), confirming specificity for the sodium channel.
Radiation inactivation yielded an estimated M(r) of 171,400 +/- 25,000 for the insect sodium channel.

Conclusions:

The cockroach nervous system possesses specific saxitoxin binding sites associated with voltage-operated sodium channels.
The estimated molecular weight provides insights into the structure of insect sodium channels.