PURIFICATION AND PROPERTIES OF ALKALINE RIBONUCLEASE FROM HUMAN-SERUM

被引:83
作者
AKAGI, K [1 ]
MURAI, K [1 ]
HIRAO, N [1 ]
YAMANAKA, M [1 ]
机构
[1] KYUSHU UNIV, FAC MED, DEPT INTERN MED 2, FUKUOKA 812, JAPAN
关键词
D O I
10.1016/0005-2787(76)90311-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Five alkaline RNase (EC 3.1.4.22) were purified .apprx. 140- to 1900-fold from human serum by phosphocellulose and DEAE-cellulose chromatographies and Sephadex G-75 filtration, with a total recovery of 22%. These were designated as RNases 1-5. Optimum activities were observed at pH 8.5-8.7 for RNases 1-4, and at pH 7.5 for RNase 5. The molecular weights of these enzymes were estimated by gel filtration as 45,000, 32,000, 20,000, 13,000 and 8500, respectively. These RNases were heat-labile proteins but are markedly stabilized with bovine plasma albumin. The reaction was activated by Na+, K+, Mg2+ and Ca2+, and inhibited by Co2+, Fe2+, Cu2+ and Zn2+. EDTA had little effect on the velocity of the reaction. Spermine caused 2- to 7-fold activation. Among the substrates examined, these RNases preferentially hydrolyzed pyrimidine bodies and except for RNase 5, had a higher affinity for poly(C) than poly(U) as substrate. Each enzyme was free from other nucleolytic enzymes and hydrolyzed only RNA.
引用
收藏
页码:368 / 378
页数:11
相关论文
共 27 条
[11]   SERUM RIBONUCLEASE ACTIVITY [J].
HOUCK, JC ;
BERMAN, LB .
JOURNAL OF APPLIED PHYSIOLOGY, 1958, 12 (03) :473-476
[12]  
LAVES W, 1952, BIOCHEM Z, V322, P292
[13]  
LEVY AL, 1960, CLIN CHEM, V6, P43
[14]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[15]  
MANSSON PE, 1974, SCAND J HAEMATOL, V12, P284
[16]  
Naskalski J, 1972, Przegl Lek, V29, P394
[17]   RIBONUCLEASES OF CHRONIC GRANULOCYTIC LEUKAEMIA LEUCOCYTES [J].
NASKALSKI, J ;
SZNAJD, J .
NATURE, 1967, 215 (5099) :414-+
[18]  
RABINOVITCH M, 1959, J LAB CLIN MED, V53, P563
[19]   INCREASE IN SERUM RIBONUCLEASE ACTIVITY AFTER BILATERAL NEPHRECTOMY [J].
RABINOVITCH, M ;
DOHI, SR .
AMERICAN JOURNAL OF PHYSIOLOGY, 1956, 187 (03) :525-528
[20]  
RABINOVITCH M, 1959, P SOC EXP BIOL MED, V100, P865