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Purification and characterization ofFus sI3596, a 65 kd allergen ofFusarium solani

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Abstract

A component ofFusarium solani (F. solani), identified as the major allergen,Fus sI3596 * was purified to homogeneity from culture filtrate (CF) by means of anion-exchange column chromatography, gel filtration and FPLC. The homogeneity ofFus sI3596 * was assessed by IEF, PAGE, SDS-PAGE (non-reducing), immunoblot and HPLC.Fus sI3596 * was isolated as a glycoprotein of MW 65 kd and pI 3.6. The IgE ELISA-inhibition assay after periodate treatment of the fraction showed a lower IgE binding capacity suggesting involvement of carbohydrate moiety in IgE binding reactions of the allergen. Peptide fragments ofFus sI3596 * obtained after CNBr and trypsin treatment were analysed by immunoblotting for their allergenicity. This study indicated that there could be at least 3 allergenic determinants in the major allergen,Fus sI3596 * ofF. solani CF.

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Abbreviations

DEAE:

diethyl aminoethyl

HPLC:

high performance liquid chromatography

SDS:

sodium dodecyl sulphate

PBS-Tween:

phosphate buffer saline containing 0.1% tween

DTT:

dithiothreitol

TFA:

triflouroacetate

FPLC:

fast protein liquid chromatography

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  1. Centre for Biochemical Technology, University Campus, Mall Road, Near Jubilee Hall, 110007, Delhi, India

    Jyotsna Verma, Santosh Pasha & Sharad V. Gangal

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  1. Jyotsna Verma
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  2. Santosh Pasha
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  3. Sharad V. Gangal
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Verma, J., Pasha, S. & Gangal, S.V. Purification and characterization ofFus sI3596, a 65 kd allergen ofFusarium solani . Mol Cell Biochem 131, 157–166 (1994). https://doi.org/10.1007/BF00925952

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  • DOI: https://doi.org/10.1007/BF00925952

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