Study on DNase activity specific from Haemonchus contortus reproductive tissue

Kwak, Dongmi

Korean Journal of Veterinary Research 2004;44(3):441-448.

Dongmi Kwak

Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University

염전위충(Haemonchus contortus) 생식기관의 DNase활성에 관한 연구

곽동미

워싱턴주립대학교 수의과대학

Abstract

DNase activity in Haemonchus contortus reproductive tissue was characterized and compared to that in whole worm. DNase activity in reproductive tissue was detected throughout pHs 4-10 with high activity under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. The activity produced DNA fragments with mixtures of 3'-hydroxyls (OH) and 3'- phosphates (P) at each pH. Three distinct DNase activities were identified and had $M_rs$ of 34, 36 and 38.5 kDa in zymograms, which were distinguished according to pH requirement and sensitivity to EDTA. Among them, the 36 kDa reproductive tissue DNase had predominant activity at pH 5.0, but very weak at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 36 kDa reproductive tissue DNase resemble those of classic acidic DNases. In contrast, 36 kDa whole worm DNase activity had high activity at both pH 5.0 and 7.0. While the 36 kDa DNase activity at pH 5.0 was similar in both reproductive tissue and whole worm samples, the activity at pH 7.0 was predominantly detected in whole worm sample. This suggests that the 36 kDa whole worm DNase at pH 5.0 differs from that at pH 7.0. Thus, results indicate that the EDTA-insensitive 36 kDa DNase at pH 5.0 is specific for H. contortus reproductive tissue.

Key Words: reproductive tissue, DNase, activity, Haemonchus contortus, parasite