PURIFICATION AND MOLECULAR-CLONING OF MOUSE RENAL DIPEPTIDASE

被引:25
作者
SATOH, S
KEIDA, Y
KONTA, Y
MAEDA, M
MATSUMOTO, Y
NIWA, M
KOHSAKA, M
机构
[1] FUJISAWA PHARMACEUT CO LTD,DEPT BIOTECHNOL,PROD DEV LABS,1-6-2 CHOME,YODOGAWA KU,OSAKA 532,JAPAN
[2] HIROSAKI UNIV,DEPT INTERNAL MED 2,HIROSAKI,AOMORI 036,JAPAN
关键词
PURIFICATION; AMINO ACID SEQUENCE; DNA CLONING; RENAL DIPEPTIDASE; EXPRESSION; NORTHERN BLOTTING; (MOUSE);
D O I
10.1016/0167-4838(93)90157-M
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
Mouse renal dipeptidase (mouseRDP, EC 3.4.13.11) was purified from the membrane fraction of kidney. The molecular mass of the enzyme was 115 kDa by size-exclusion HPLC and SDS-PAGE under non-reduced conditions and 58 kDa by SDS-PAGE under reduced conditions. The mouseRDP cDNA fragment was amplified from mouse kidney total RNA by reverse transcriptation-polymerase chain reaction (RT-PCR). The mouseRDP cDNA was isolated from a kidney cDNA library using the probe. The primary structure of mouseRDP deduced from the cDNA showed a high homology with renal dipeptidase from various mammals, except for the amino-terminal and carboxy-terminal domains. Recombinant mouseRDP obtained from transfected mouse L929 cells containing the expression plasmids has the same K(m) value and molecular mass as native mouse renal dipeptidase. From Northern blotting analysis, expression of the mouseRDP gene was recognized in both kidney and liver.
引用
收藏
页码:234 / 242
页数:9
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