Localization of Cytosolic NADP-dependent Isocitrate Dehydrogenase in the Peroxisomes of Rat Liver Cells
Localization of Cytosolic NADP-dependent Isocitrate Dehydrogenase in the Peroxisomes of Rat Liver Cells
复制标题
大鼠肝细胞过氧化物酶体中胞质 NADP 依赖性异柠檬酸脱氢酶的定位
DOI：
--
复制
发表时间：
2001
影响因子：
3.2
通讯作者：
S. Yokota
中科院分区：
文献类型：
--
作者：
Takashi Yoshihara;T. Hamamoto;Ryo Munakata;Ryosuke Tajiri;M. Ohsumi;S. Yokota
Two types of NADP-dependent isocitrate dehydrogenases (ICDs) have been reported: mitochondrial (ICD1) and cytosolic (ICD2). The C-terminal amino acid sequence of ICD2 has a tripeptide peroxisome targeting signal 1 sequence (PTS1). After differential centrifugation of the postnuclear fraction of rat liver homogenate, approximately 75% of ICD activity was found in the cytosolic fraction. To elucidate the true localization of ICD2 in rat hepatocytes, we analyzed the distribution of ICD activity and immunoreactivity in fractions isolated by Nycodenz gradient centrifugation and immunocytochemical localization of ICD2 antigenic sites in the cells. On Nycodenz gradient centrifugation of the light mitochondrial fraction, ICD2 activity was distributed in the fractions in which activity of catalase, a peroxisomal marker, was also detected, but a low level of activity was also detected in the fractions containing activity for succinate cytochrome C reductase (a mitochondrial marker) and acid phosphatase (a lysosomal marker). We have purified ICD2 from rat liver homogenate and raised a specific antibody to the enzyme. On SDS-PAGE, a single band with a molecular mass of 47 kD was observed, and on immunoblotting analysis of rat liver homogenate a single signal was detected. Double staining of catalase and ICD2 in rat liver revealed co-localization of both enzymes in the same cytoplasmic granules. Immunoelectron microscopy revealed gold particles with antigenic sites of ICD2 present mainly in peroxisomes. The results clearly indicated that ICD2 is a peroxisomal enzyme in rat hepatocytes. ICD2 has been regarded as a cytosolic enzyme, probably because the enzyme easily leaks out of peroxisomes during homogenization. (J Histochem Cytochem 49:1123–1131, 2001)
编码猪线粒体 NADP 特异性异柠檬酸脱氢酶的 cDNA 的分离和序列。
影响因子：
2.9
作者：
Haselbeck,RJ;Colman,RF;McAlister-Henn,L
通讯作者：
McAlister-Henn,L
法尼基二磷酸合酶位于过氧化物酶体中。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Krisans,SK;Ericsson,J;Edwards,PA;Keller,GA
通讯作者：
Keller,GA
胞质 NADP( ) 依赖性异柠檬酸脱氢酶。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Jennings,GT;Sechi,S;Stevenson,PM;Tuckey,RC;Parmelee,D;McAlister-Henn,L
通讯作者：
McAlister-Henn,L