Article

Structure and function of a phospholipid hydroperoxide glutathione peroxidase-like protein from barley

Laboratory of Biochemistry, Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, Okayama 710-0046, Japan; Barley Germplasm Center, Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, Okayama 710-0046, Japan
Journal of Molecular Catalysis B: Enzymatic DOI:10.1016/S1381-1177(03)00104-8 pp.397-403

ABSTRACT A cDNA encoding barley phospholipid hydroperoxide glutathione peroxidase (PHGPX)-like protein was cloned and sequenced by the reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods. The cDNA comprised 846 bp, and included an open reading frame which encodes a polypeptide of 169 amino acid residues with a molecular mass of 18,532 Da. The deduced amino acid sequence showed significant identity to plant putative PHGPXs and mammalian PHGPXs. The cloned gene was expressed in Escherichia coli cells to produce an extra protein, which showed a molecular mass similar to the deduced one, and the clone cells were much more tolerant to NaCl stress than the host cells.

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    Article: Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx).
    Li-guang Shi, Wen-juan Xun, Wen-bin Yue, Chun-xiang Zhang, You-she Ren, Qian Wang, Xiao-ying Wu, Lei Shi, Ru-jie Yang, Fu-lin Lei
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    ABSTRACT: Phospholipid hydroperoxide glutathione peroxidase (PHGPx), as a ubiquitous antioxidant enzyme in the glutathione peroxidases (GPx) family, plays multiple roles in organisms. However, there is very little information on PHGPx in goats (Capra hircus). In this study, a full-length cDNA was cloned and characterized from Taihang black goat testes. The 844 bp cDNA contains an open reading frame (ORF) of 597 bp. The goat PHGPx nucleotide sequence contains a selenocysteine (sec) codon TGA(244-246), two potential start codons ATG(20-22) and ATG(108-110), a polyadenylation signal AATAAA(813-818) and selenocysteine insertion sequence (SECIS) motif AUGA(688-691), UGA(729-731) and AAA(703-705). As a selenoprotein, the active-site motifs and GPx family signature motifs LAFPCNQF(101-108) and WNFEK(165-170) were also found. The order of PHGPx mRNA expression levels was: testes > heart > brain > epididymis > kidney > liver > lung > spleen > muscle. Real-time PCR and immunohistochemistry results revealed similar expression differences in different age testes, with high expression levels during adolescence. Immunofluorescence results suggested that PHGPx mainly expressed in Leydig cells and spermatids in mature goat testes.
    International journal of biological sciences 01/2010; 6(4):316-26. · 2.70 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

169 amino acid residues
 
cDNA encoding barley phospholipid hydroperoxide glutathione peroxidase
 
clone cells
 
deduced amino acid sequence
 
Escherichia coli cells
 
host cells
 
molecular mass
 
open reading frame
 
plant putative PHGPXs
 
rapid amplification
 
reverse transcription-polymerase chain reaction
 
sequenced
 
tolerant
 

Manabu Sugimoto