Effect of Tunicamycin and 2-deoxy-D-glucose on the Cellular and Secreted Isoperoxidase Levels of Tobacco Callus

국문 초록 (Abstract)

담배 callus 배양시 tunicamycin(TM)과 2-deoxy-D-glucose(2-D-G)가 세포내 peroxidase와 배지로 분비되는 peroxidase의 활성과 isozyme pattern에 미치는 영향을 조사하였다. TM과 2-D-G 처리에 의하여 세포내 peroxidase의 비활성도는 전 배양기간동안 현저히 증가한 반면 배지로 분비되는 peroxidase의 비활성도는 배양초기를 제외하고는 점차 감소하였다. Isozyme patterns변화를 조사한 결과 M과 2-D-G처리에 의하여 세포내 peroxidase 중 glycoprotein인 cathodic isoperoxidase $C_4$ 가 뚜렷한 증가를 보인 반면 또 다른 glycoprotein인 anodic isoperoxidase $A_3$는 감소를 나타냈다. 한편 TM과 2-D-G 처리시 배지로 분비되는 peroxidase의 비활성도 감소는 주로 isoperoxidase $A_3$의 감소에 기인한다는 사실도 확인하였다. Anti-$A_3$ 혈청과 anti-$C_4$혈청을 사용하여 isoperoxidase $C_4$와 $A_3$의 양의 변화를 정량한 결과 비활성도 변화 및 pattern 변화와 일치하는 것으로 나타났다.

다국어 초록 (Multilingual Abstract)

The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase $C_4$ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase $A_3$ which is also a glycoprotein. Immunoprecipitation with anti-$A_3$ antibody and anti-$C_4$ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular $C_4$ activity was due to the increased amount of $C_4$ proteins and the decrease of the activity of secretory isoperoxidase $A_3$ was caused by the decrease of $A_3$ proteins in the medium in the presence of the glycosylation inhibitors.