Homogeneity of Phospholipase C of Bovine Uterus and Seminal Vesicle Compared with Brain Isozymes

소의 자궁 및 고환에서 Phospholipase C의 분리 및 뇌 Isozyme과의 비교 연구

  • Kim, Jung-Hye (Department of Biochemistry, College of Medicine, Yeungnam University) ;
  • Rhee, Sue-Goo (Department of Biochemistry, National Institute of Health) ;
  • Lee, Ki-Yung (Department of Biochemistry, College of Medicine, Yeungnam University)
  • 김정희 (영남대학교 의과대학 생화학교실) ;
  • ;
  • 이기녕 (영남대학교 의과대학 생화학교실)
  • Published : 1988.12.30

Abstract

Phosphoinositide-specific phospholipase C(PI-PLC) is a second messenger of signal transducer on cell membrane. In the previous study, PLC of bovine brain has been purified three isozymes. In this paper, uterus and seminal vesicle have been purified. Two peaks of PI-PLC activity were resolved when bovine uterus and seminal vesicle proteins were chromatographed on a DEAE and phenyl TSK 5PW HPLC column. Each two peak was compared with PI-PLC I, IT and ill from bovine brain and we got the retension time on HPLC. The peak fractions with PLC activity were tested homogeneity with brain PLC monoclonal antibodies(Mab). Mab-labeled affigels were bounded in the range of 73.8%~97.5% with PLC I, IT and III. Homogeneity of fractions were revealed that DEAE F-1 and phenyl F-1-I were highest level of PLC III in uterus and seminal vesicle and DEAE F-2 and phenyl F-2-I were mixed PLC I and II.

Phospholipase C는 second messenger로서 세포 밖의 signal transduction에 중요한 효소 이다. 본 연구에서 소의 뇌, 자궁 및 고환에서 high performance liquid chromatography(HPLC)를 이용하여 phospholipase C(PLC)를 추출하였으며 뇌에서 3개의 isozyme(F-1, F-2, F-3), 자궁과 고환에서 각각 2개의 isozyme을 얻었고 HPLC에서의 retension time을 구하였다. Homogeneity 검사를 위하여 소 brain의 각 isozyme I, II 및 III에 대한 PLC-monoclonal antibody(Mab)를 affigel에 label 시켰고 결합능(binding capacity)는 73.8~97.5%였다. PLC-Mab와 자궁 및 고환의 PLC isozyme과의 homogeneity 검사에서 binding capacity는 자궁의 F-1은 PLC III가 주이며 F-2는 DEAE column에서는 II가 주이나 phenyl column에선 I과 II가 주이고 고환에선 F-1은 PLC III가 주이고 F-2에서는 PLC II가 주인 것으로 나타났다.

Keywords