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Korean J Parasitol > Volume 29(2):1991 > Article

Original Article
Korean J Parasitol. 1991 Jun;29(2):113-120. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1991.29.2.113
Copyright © 1991 by The Korean Society for Parasitology
Molecular weight of major component proteins in crude saline extract of adult Paragonimus westermani
Y Kong,WB Kim,SY Kang and SY Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract

When the component proteins in crude saline extract of 13-week old adult Paragonimus westermani were observed by non-denaturing discontinuous-polyacrylamide gel electrophoresis (Disc-PAGE), 8 distinct bands were clearly recognized. Molecular weight (MW) of each band protein, numbered in sequence from cathodal side which appeared in 10% separating gel, was measured first by Ferguson plot utilizing different gel concentrations from 10% to 4.5%. MW of band 1 protein (known as egg protein) was 440 kDa. And MW of other band proteins were: 386 kDa in band 2, 17.4 kDa in band 3, 17 kDa in band 4, 14.3 kDa in band 5, 46 kDa in band 6, 38 kDa in band 7 and 23 kDa in band 8. When the proteins in the crude extract were separated into fractions by molecular sieve chromatography through 1.6(φ) × 70 cm sized Sephacryl S-300 Superfine column and revisualized by Disc-PAGE in 8% gel, the sequence of eluted proteins was band 1, band 2, band 6, band 7 and bands 3, 4, 5 and 8. This elution profile confirmed MW of each band protein in the crude extract as measured by Ferguson plot.

Figures


Fig. 1
Separation of component proteins in crude saline extract of adult P. westermani at different gel concentrations in Disc-PAGE. A: 10% separationg gel, B: 9% gel, C: 8% gel, D: 7% gel, E: 6% gel, F: 5.5% gel, G:5% gel and H: 4.5% gel. Protein bands were numbered as appeared from cathodal side in 10% separation gel.


Fig. 2
Calculation of minus slope of 100·log(Rƒ·100) against % gel concentration of each protein band in the saline extract of P. westermani. Numericals stand form the band number in Fig. 1.


Fig. 3
Calibration of molecular weight of the component proteins in the crude saline extract from the minus slopes. Numericals mean the band number in Fig. 1.


Fig. 4
Elution profile of crude extract of P. westermani. filtrated through Sephacryl S-300 Superfine. Eluent: 0.15M PBS containing 0.01% merthiolate and 0.006% PMSF. Flow rate: 15ml·cm-2·hr. Collection volume: 35 drops/tube.


Fig. 5
Banding patterns of each fraction collected by Sephacryl S-300 Superfine. Proteins in each fraction wee revisualized in 8% Disc-PAGE. C: Crude saline extract, 1: Fraction 1, 2:Fraction 2, 3: Fraction 3, 4: Fraction 4, 5: Fraction 5, 6: Fraction 6.

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