Identification and characterization of human PMP34, a protein closely related to the peroxisomal integral membrane protein PMP47 of Candida boidinii.
Identification and characterization of human PMP34, a protein closely related to the peroxisomal integral membrane protein PMP47 of Candida boidinii.
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人 PMP34 的鉴定和表征，该蛋白与博伊丁假丝酵母的过氧化物酶体整合膜蛋白 PMP47 密切相关。
DOI：
10.1046/j.1432-1327.1998.2580332.x
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
P. Veldhoven
中科院分区：
文献类型：
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作者：
T. Wylin;M. Baes;C. Brees;G. Mannaerts;M. Fransen;P. Veldhoven
克隆了与博伊丁假丝酵母过氧化物酶体整合膜蛋白PMP 47（CbPMP 47）具有33%同源性的人蛋白质的cDNA。它编码一个307个氨基酸的蛋白质，计算分子量为34,566 Da，我们命名为HsPMP 34，含有六个膜跨度，与PMP 47一样，属于线粒体溶质载体家族。基于重叠的可用表达序列标签（EST）结合一个EST的测序，还可以推断小鼠对应物（分子量为34，422 Da）的氨基酸序列。通过逆转录PCR方法，发现人肝脏和小鼠、大鼠肝脏及其他组织中存在PMP 34 mRNA。人类基因定位于染色体22 q13。由于HsPMP 34不仅与CbPMP 47相似，而且与已知或推定的线粒体转运蛋白相似，因此研究了其亚细胞定位。用编码HsPMP 34/绿色荧光融合蛋白的质粒转染的HepG 2细胞或小鼠成纤维细胞的荧光显微镜显示点状图案，使人联想到过氧化物酶体分布。通过间接免疫荧光，HsPMP 34/绿色荧光融合蛋白清楚地与二聚过氧化物酶体硫解酶共定位。在转染由于PEX 5的敲除而缺乏功能性过氧化物酶体的小鼠成纤维细胞后[Baes，M.，Gressens，P.，Baumgart，E.，Carmeliet，P.，Casteels，M.，Fransen，M.，Evrard，P.，Fahimi，D.，Declercq，P.E.，Collen，D.，货车费尔德霍芬，P. P. P.（1997）Nat.Genet. 17，49-53]，荧光与较大且丰度较低的结构相关。综上所述，这些数据表明，HsPMP 34是一种过氧化物酶体膜蛋白，并且是CbPMP 47的人类对应物或与其密切相关。
The cDNA of a human protein displaying 33% identity to the peroxisomal integral membrane protein PMP47 of Candida boidinii (CbPMP47) was cloned. It encodes a protein of 307 amino acids, with a calculated molecular mass of 34,566 Da. The protein, which we name HsPMP34, contains six membrane spans and belongs, like PMP47, to the family of mitochondrial solute carriers. Based on overlapping available expressed sequence tags (ESTs) combined with the sequencing of one EST, the amino acid sequence of the mouse counterpart (molecular mass of 34,422 Da) could also be deduced. By means of reverse-transcription PCR, the presence of PMP34 mRNA was revealed in liver of man and in liver and other tissues of mouse and rat. The human gene is localized to chromosome 22q13. Given the similarity of HsPMP34 not only to CbPMP47 but also to known or putative mitochondrial transporters, its subcellular localization was investigated. Fluorescence microscopy of HepG2 cells or mouse fibroblasts transfected with a plasmid encoding a HsPMP34/green fluorescent fusion protein revealed a punctate pattern, reminiscent of a peroxisomal distribution. By indirect immunofluorescence, the HsPMP34/green fluorescent fusion protein clearly colocalized with the dimeric peroxisomal thiolase. Upon transfection of mouse fibroblasts lacking functional peroxisomes due the knock-out of PEX5 [Baes, M., Gressens, P., Baumgart, E., Carmeliet, P., Casteels, M., Fransen, M., Evrard, P., Fahimi, D., Declercq, P. E., Collen, D., Van Veldhoven, P. P. & Mannaerts, G. P. (1997) Nat. Genet. 17, 49-53], fluorescence was associated with larger and less-abundant structures. Taken together, the data indicate that HsPMP34 is a peroxisomal membrane protein and is either the human counterpart of CbPMP47 or closely related to it. According to its structure, the protein is most probably involved in transport.