Process of Denaturation of Arachin by Heat Treatment

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When arachin (S20, w, 9S) was heated in 0.01 m sodium phosphate buffer, pH 7.9, at 70°C, the 9S protein decreased, accompanied by an increase of the 3S component and precipitate. The 3S component was not precipitated by prolonged heating. The precipitate contained six arachin subunits (S1 to S6) in a molar ratio of 3/4:3/4:3/4:1:1:1, but the 3S component comprised S1, S2 and S3 as dissociated forms in an equimolar ratio. The molar ratio of S1, S2 and S3 in the 3S component to those in the precipitate was 1:3 at each time of heating. On the other hand, arachin was neither dissociated nor precipitated by heating in 0.3 M sodium phosphate buffer, pH 7.9. The fluorescence intensity of 1-anilino-8-naphtalene sulfonate (ANS) conjugated with the heated arachin in 0.3 M buffer, however, was stronger than that of ANS conjugated with the intact arachin. These results suggest that two different pathways leading to insolubilization are involved in the thermal denaturation of arachin.
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