IDENTIFICATION OF 2 ESSENTIAL HISTIDINE-RESIDUES OF RIBONUCLEASE-T2 FROM ASPERGILLUS-ORYZAE

被引:80
作者
KAWATA, Y [1 ]
SAKIYAMA, F [1 ]
HAYASHI, F [1 ]
KYOGOKU, Y [1 ]
机构
[1] OSAKA UNIV,INST PROT RES,YAMADA OKA,SUITA,OSAKA 565,JAPAN
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1990年 / 187卷 / 01期
关键词
D O I
10.1111/j.1432-1033.1990.tb15303.x
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
Ribonuclease (RNase) T2 from Aspergillus oryzae was modified by diethyl pyrocarbonate and iodoacetic acid. RNase T2 was rapidly inactivated by diethyl pyrocarbonate above pH 6.0 and by incorporation of a carboxymethyl group. No inactivation occurred in the presence of 3′AMP. 1H‐NMR titration and photo‐chemically induced dynamic nuclear polarization experiments demonstrated that two histidine residues were involved in the active site of RNase T2. Furthermore, analysis of inactive carboxymethylated RNase T2 showed that both His53 and His115 were partially modified to yield a total of one mole of Nτ‐carboxymethylhistidine/mole enzyme. The results indicate that the two histidine residues in the active site of RNase T2 are essential for catalysis and that modification of either His53 or His115 inactivates the enzyme. Copyright © 1990, Wiley Blackwell. All rights reserved
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页码:255 / 262
页数:8
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