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A Gel-Chromatographic and Light Scattering Study of the Salmonella typhi Endotoxin

Gj. Deželić ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia
N. Deželić ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia
D. Jušić ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia
B. Pende ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia
D. Sinković ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia
M. Zebec ; Laboratory of Biochemistry, Andrija Stampar School of Public Health, Faculty of Medicine, University of Zagreb, and Institute of Immunology, Zagreb, Croatia, Yugoslavia

Sažetak

The endotoxin of Salmonella typhi, strain 0-901, was isolated
by extraction with hypertonic (1 M) sodium chloride solutions and
studied by gel-chromatography and light scattering methods. The
gel-chromatographic separation was performed on Sepharose 2B,
Sepharose 4B and Sephadex G-200 gels, and the fractionated material
was monitored by ultraviolet and phenol-sulfuric acid colorimetry
as well as by a photometric latex agglutination test. The
extracted material consisted of two components: one was the high
molecular weight endotoxin and the other a protein-polysaccharide
complex of a molecular weight lower than 66,000 dalton. The light
scattering experiments of endotoxin extracts showed the average
molecular weights from 1.9 to 4.9 million dalton. The separation
of the low molecular weight proteinic component was attempted by
thermal denaturation, but this had to be abandoned owing to the
denaturation and degradation of endotoxin. A high molecular weight
endotoxin component was isolated by elution on a Sephadex G-200
column and had the molecular weight of 5.6 million dalton, which
was in good accord with the value previously determined for a
Boivin extraction sample. The high molecular weight endotoxin
sample proved to be a highly polydispersed material. From the
estimates of various averages of gyration radii it has been concluded
that the particles of this sample have a more compact structurE
than those of the Boivin extraction sample, possibly due to the
tertiary structuring of polypeptide chains in the protein-lipopolysaccharide
complex of the endotoxin particle.

Ključne riječi

Hrčak ID:

196431

URI

https://hrcak.srce.hr/196431

Datum izdavanja:

25.3.1977.

Posjeta: 438 *