Microheterogeneity

There are different glycosylated proteins in snake venoms, but no glycosylated representatives of a large family of three-fingered toxins have previously been detected. A new glycoprotein was isolated from the venom of the Thai cobra Naja kaouthia. MALDI MS of the glycoprotein contained an array of peaks in the range from
8900 to
9400 Da indicating its microheterogeneity. Carbohy-drate analysis showed the presence of mannose, galactose, N-acetylglucosamine, fucose and neuraminic acid.

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The catalytic module ofHypocrea jecorina(previously Trichoderma reesei) Cel7B was homologously expressed by transformation of strain QM9414. Post-translational modi-fications in purified Cel7B preparations were analysed by enzymatic digestions, high performance chromatography, mass spectrometry and site-directed mutagenesis. Of the five potential sites found in the wild-type enzyme, only Asn56 and Asn182 were found to be N-glycosylated.

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