RAT HEMOPEXIN - MOLECULAR-CLONING, PRIMARY STRUCTURAL CHARACTERIZATION, AND ANALYSIS OF GENE-EXPRESSION

被引:56
|
作者
NIKKILA, H
GITLIN, JD
MULLEREBERHARD, U
机构
[1] CORNELL UNIV, MED CTR, COLL MED, DEPT PHARMACOL, NEW YORK, NY 10021 USA
[2] CORNELL UNIV, MED CTR, COLL MED, DEPT BIOCHEM, NEW YORK, NY 10021 USA
[3] WASHINGTON UNIV, SCH MED, DEPT PEDIAT, ST LOUIS, MO 63110 USA
关键词
D O I
10.1021/bi00217a036
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
A full-length hemopexin cDNA was isolated from a rat liver cDNA library and the derived amino acid sequence was obtained. Rat hemopexin shows a 76% amino acid homology with human hemopexin. The amino-terminal domain of rat hemopexin contains two histidine residues that are conserved in the human and rat sequences and are the most likely heme axial ligands. Analogous to human hemopexin, the rat hemopexin consists of 10 internal repeating peptide motifs characteristic of the pexin gene family. A complete conservation of cysteine residues is seen between the human and rat sequence suggesting an identical disulfide bridge structure in both proteins. Our analysis of the primary structure of rat hemopexin reveals characteristics typical for members of the pexin gene family and suggests a conserved evolutionary role for the C-terminal (non-heme-binding) domain of this protein. The full-length rat hemopexin cDNA was used to analyze changes in hemopexin gene expression during development and experimental inflammation. RNA blot analysis showed a single 2.0-kb hemopexin mRNA present in fetal liver at day 14. Hemopexin-specific mRNA was not detected in embryonic or fetal tissues at earlier stages of development and was confined to the liver throughout fetal, newborn, and adult life. The abundance of hemopexin mRNA was found to increase throughout gestation, with a sharp increase in the first postnatal weeks, reaching maximum levels in adult animals. Endotoxin-induced inflammation resulted in a 5-fold increase in hepatic hemopexin mRNA content within 48 h without associated changes in hemopexin transcript size. Adult animals exposed to hyperoxia (95% oxygen) showed a 3-fold increase in hepatic hemopexin mRNA content. Extrahepatic hemopexin gene expression was not detected during inflammation or hyperoxic exposure. The data also indicate that typical of the acute-phase response in rodents the regulation of hemopexin synthesis occurs at a pretranslational level. In contrast to many plasma proteins, hemopexin gene expression is entirely confined to the liver during development, inflammation, and tissue injury.
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收藏
页码:823 / 829
页数:7
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