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Korean J Parasitol > Volume 16(1):1978 > Article

Original Article
Korean J Parasitol. 1978 Jun;16(1):26-40. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1978.16.1.26
Copyright © 1978 by The Korean Society for Parasitology
Study on the chromosomal proteins of Fasciola hepatica
Soon-Hyung Lee,Chul-Yong Song,Keun-Bae Lee and Hi-Sung Lee
Department of Parasitology, College of Medicine, Chung-Ang University, Korea.
Department of Biochemistry, College of Medicine, Chung-Ang University, Korea.
Abstract

In attempt to investigate histone fractions and non-histones of parasites, nuclei were isolated from Fasciola hepatica by the procedure of Pogo et al. (1966). Histone fractions H1, H2a, H2b, H3 and H4 were prepared from isolated nuclei by the procedure of Johns (1964 and l967). The five histone fractions found in most tissues were also present in the Fasciola hepatica histones. These histone fractions were characterized by amino acid analysis and by polyacrylamide disc gel electrophoresis. Non-histone proteins were extracted from isolated Fasciola hepatica nuclei and separated by SDS-polyacrylamide gel electrophoresis.

The results of the experiment were summarized as follows:

1. The yield of whole histone recovered was 2.47 mg per 1 g of Fasciola hepatica.

2. The yield of DNA was 1.02 mg per gm of tissues. Consequently the DNA to histone ratio was 1:2.44.

3. The relative amounts of five fractions, i.e., Hl, H2a, H2b, H3 and H4 were 19.96%, 26.48%, 29.60%, 12.56% and 14.37%, respectively.

4. Amino acid analysis of the individual histone fractions showed that the over-all compositions were similar but not identical to those of corresponding fraction from calf thymus.

5. It was found that histone H2b fraction of Fasciola hepatica contained detectable amounts of ε-N-monomethyllysine. No evidence for the presence of methylated lysine or other side-chain derivatives was reported on this histone fraction.

6. In SDS-polyacrylamide disc gel, it showed that 17 protein bands of nuclear acidic protein can be identified visually.

Figures


Fig. 1
Chromatographic separation of the amino acids of Fasciola hepatica histone fraction H1 under the standard conditions described under "Materials and Methods". Ninhydrin color intensity is plotted against the time of elution.


Fig. 2
Elution profile of a standard amino acid mixture containing the methylated derivatives of lysine and histidine. This standard amino-acid mixture was separated on the amine analyzer column under the conditions described under "Materials and Methods". Nin-hydrin color intensity is plotted against the time of elution. DML: N-dimethyllysine; MML: N-monomethyllysine; 3-MH: 3-methy-1histidine.


Fig. 3
Chromatography of the amino acids of Fasciola hepatica histone fraction H2b under the conditions described in Fig. 2. Ninhydrin color intensity is plotted against the time of elution, beginning with the appearance of the lysine peak.


Fig. 4
Non-histone proteins which were isolated from Fasciola hepatica were soluble in 0.01 M sodium phosphate buffer(pH 7.2) containing 0.1% sodium dodecy1 sulphate and 0.14M 2-mercaptoethanol. Samples containing 200µg of proteins were analyzed by 10% polyacry-lamide-gel electrophoresis at room temperature, at a constant voltage of 6V/cm. The gels were stained with Amido Black 1OB.


Fig. 5
Non-histone proteins were obtained from Fasciola hepatica nuclei. These proteins were fractionated by polyacrylamide gel electrophoresis. The gels were stained with Amido Black 10B and scanned at 570 nm as described in the "Materials and Methods".

Tables


Table I
Contents of DNA and total histone from Fasciola hepatica


Table II
Yields of histone from Fasciola hepatica


Table III
Relative yields of histone fractions from various sourses. Values are expressed as per cent of total histone


Table IV
Amino acid compostition of histone fractions from Fasciola hepatica. Results are given as moles/100 moles of all amino acids recovered


Table V-1
Amino acid compositions of histones from Fasciola hepatica in comparison with those of calf thymus


Tables V-2
Amino acid compositions of histones from Fasciola hepatica comparison with those of calf thymus


Table VI
Contents of methylated amino acid residues in histone fractions of Fasciola hepatica. Values are expressed as moles/100 moles of all amino acids recovered.


Table VII
Distribution of protein in extracts of Fasciola hepatica nuclei

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