As a preliminary study for the explanation of pathobiology of Neodiplostomum seoulense infection, a 54 kDa protease was purified from the crude extract of adult worms by sequential chromatographic methods. The crude extract was subjected to DEAE-Sepharose Fast Flow column, and protein was eluted using 25 mM Tris-HCl (pH 7.4) containing 0.05, 0.1, 0.2 and 0.4 M NaCl in stepwise elution. The 0.2 M NaCl fraction was further purified by Q-Sepharose chromatography and protein was eluted using 20 mM sodium acetate (pH 6.4) containing 0.05, 0.1, 0.2 and 0.3 M NaCl, respectively. The 0.1M NaCl fraction showed a single protein band on SDS-PAGE carried out on a 7.5-15% gradient gel. The proteolytic activities of the purified enzyme were specifically inhibited by L-trans-epoxy-succinylleucylamide (4-guanidino) butane (E-64) and iodoacetic acid. The enzyme, cysteine protease, showed the maximum proteolytic activity at pH 6.0 in 0.1 M buffer, and degraded extracellular matrix proteins such as collagen and fibronectin with different activities. It is suggested that the cysteine protease may play a role in the nutrient uptake of N. seoulense from the host intestine.
Citations
Citations to this article as recorded by
Roles of Cysteine Proteases in Biology and Pathogenesis of Parasites Nawu Yang, Maurice A. Matthew, Chaoqun Yao Microorganisms.2023; 11(6): 1397. CrossRef
Foodborne Intestinal Flukes in Southeast Asia Jong-Yil Chai, Eun-Hee Shin, Soon-Hyung Lee, Han-Jong Rim The Korean Journal of Parasitology.2009; 47(Suppl): S69. CrossRef
Antigenic Properties of Cystatin-binding Cysteine Proteinases from Neodiplostomum seoulense Tae-Yun Kim, Kye-Young Han, Eun-Hee Shin, Jong-Yil Chai Journal of Parasitology.2008; 94(3): 654. CrossRef
Antibody Responses in Sera of Different Mouse Strains Experimentally Infected with Neodiplostomum seoulense Eun-Taek Han, Jun-Hu Chen, Jong-Yil Chai The Korean Journal of Parasitology.2008; 46(4): 279. CrossRef
Antigenic Properties of Cystatin-binding Cysteine Proteinases from Neodiplostomum seoulense Tae-Yun Kim, Kye-Young Han, Eun-Hee Shin, Jong-Yil Chai Journal of Parasitology.2008; 94(3): 654. CrossRef
Food-borne intestinal trematode infections in the Republic of Korea Jong-Yil Chai, Soon-Hyung Lee Parasitology International.2002; 51(2): 129. CrossRef
Cystatin Capture Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Human Clonorchiasis and Profile of Captured Antigenic Protein ofClonorchis sinensis Tae Yun Kim, Shin-Yong Kang, Sun Hyo Park, Kom Sukontason, Kabkaew Sukontason, Sung-Jong Hong Clinical Diagnostic Laboratory Immunology.2001; 8(6): 1076. CrossRef
