Signal peptide of eosinophil cationic protein is toxic to cells lacking signal peptide peptidase

Eosinophil cationic protein (ECP) is a toxin secreted by activated human eosinophils. The properties of mature ECP have been well studied but those of the signal peptide of ECP (ECPsp) are not clear. In this study, several chimeric proteins containing N-terminal fusion of ECPsp were generated, and i...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2004-09, Vol.322 (2), p.585-592
Main Authors: Wu, Chia-Mao, Chang, Margaret Dah-Tsyr
Format: Article
Language:English
Subjects:
Aspartic Acid Endopeptidases - deficiency
Aspartic Acid Endopeptidases - genetics
Aspartic Acid Endopeptidases - metabolism
Blood Proteins - genetics
Blood Proteins - metabolism
Blood Proteins - toxicity
Eosinophil cationic protein
Eosinophil Granule Proteins
Escherichia coli
Genes, Reporter
Growth inhibition
Human signal peptide peptidase
Humans
Pichia
Pichia pastoris
Protein Sorting Signals - genetics
Protein Sorting Signals - physiology
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Ribonucleases - genetics
Ribonucleases - metabolism
Ribonucleases - toxicity
RNA, Messenger - metabolism
Signal peptide
siRNA
Toxic signal peptide
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Summary:Eosinophil cationic protein (ECP) is a toxin secreted by activated human eosinophils. The properties of mature ECP have been well studied but those of the signal peptide of ECP (ECPsp) are not clear. In this study, several chimeric proteins containing N-terminal fusion of ECPsp were generated, and introduced into Escherichia coli, Pichia pastoris, and human epidermoid carcinoma cell line A431 to study the function of ECPsp. We found that expression of ECPsp chimeric proteins inhibited the growth of E. coli and P. pastoris but not A431 cells. Primary sequence analysis and in vitro transcription/translation of ECPsp have revealed that it is a potential substrate for human signal peptide peptidase (hSPP), an intramembrane protease located in endoplasmic reticulum. In addition, knockdown of the hSPP mRNA expression in ECPsp-eGFP/A431 cells caused the growth inhibitory effect, whereas complementally expression of hSPP in P. pastoris system rescued the cell growth. Taken together, we have demonstrated that ECPsp is a toxic signal peptide, and expression of hSPP protects the cells from growth inhibition.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2004.07.160