Korean J Parasitol.  1992 Sep;30(3):191-199. 10.3347/kjp.1992.30.3.191.

Isolation and partial characterization of cysteine proteinase from sparganum

• Song CY
• Choi DH
• Kim TS
• Lee SH

• KMID: 1715521
• DOI: http://doi.org/10.3347/kjp.1992.30.3.191

Abstract

A proteolytic enzyme was purified from the tissue extract of spargana (plerocercoids of Spirometra erinacei) by DEAE-Trisacryl M ion exchange chromatography and thiopropyl-sepharose affinity chromatography resulted in a 21-fold purification. The proteinase activity was assayed with a synthetic fluorescent substrate, carbobenzoxy-phenylalanyl-7-amino-4-trifluoromethyl-coumarin. SDS-polyacrylamide gel electrophoresis of the purified materials revealed a single 28,000 dalton band. Inhibitor profiles of the band indicated that it belonged to cysteine endopeptidases. It exhibited identical pH curves with optimum at pH 5.5, and 50% activity from pH 4.7 to 8. It could completely degrade collagen chains to three identical products. It also showed some activity on hemoglobin. Furthermore, the band on immunoblots was reactive to the sera of sparganosis patients. These results suggest that the proteolytic enzyme belongs to cysteine proteinase which plays a role in the tissue penetration. Also it may be used as the antigen for diagnosis of active sparganosis.

MeSH Terms

Chromatography,-Affinity
Electrophoresis,-Polyacrylamide-Gel
Hydrogen-Ion-Concentration
Immunoblotting-
*Cysteine-Proteinases-isolation-and-purification
*Sparganum-enzymology
Cysteine-Proteinases