2003年5月
Heterologous expression of Ascaris suum cytochrome b(5) precursor protein: a histidine-tagged full-length presequence is correctly processed to transport the mature protein to the periplasm of Escherichia coli
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
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- 巻
- 413
- 号
- 2
- 開始ページ
- 253
- 終了ページ
- 261
- 記述言語
- 英語
- 掲載種別
- DOI
- 10.1016/S0003-9861(03)00124-3
- 出版者・発行元
- ACADEMIC PRESS INC ELSEVIER SCIENCE
The cytochrome b(5) of the body wall of adult Ascaris suum, a porcine parasitic nematode, is a novel type of cytochrome b(5). It is a soluble protein that lacks the COOH-terminal membrane-anchoring domain found in erythrocyte cytochrome b5, but possesses an NH2-terminal extension (presequence) of 30 amino acids that are missing from the 82-residue protein purified from the nematode tissues [Yu, Y., Yamasaki, H., Kita, K., and Takamiya, S., 1996, Arch. Biochem. Biophys. 328, 165-172]. The nematode cytochrome b5 is, therefore, probably synthesized as a precursor protein whose presequence is cleaved to form a mature protein, but the localization of the mature protein is still unknown. To investigate the processing of the putative precursor protein, the wild-type precursor of nematode cytochrome b5 with a complete presequence (b5wt) and its NH2 terminus-truncated derivatives, b5Delta18 and b5Delta28, with 18 and 28 residues deleted. respectively, were expressed using pET-28a(+) vector in Escherichia coli. As expected, all transformants, tb5wt, tb5Delta18, and tb5Delta28, produced recombinant proteins with a histidine-tagged NH2-terminal extension. However, only the recombinant protein with the full-length presequence, produced in tb5wt, was correctly processed and transported to the periplasm, from which the majority of the induced product was purified as a mature protein chemically and functionally identical to the native protein purified from the nematode body wall. These results clearly show that the nematode histidine-tagged presequence functions as a signal peptide in E. coli. (C) 2003 Elsevier Science (USA). All rights reserved.
- リンク情報
- ID情報
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- DOI : 10.1016/S0003-9861(03)00124-3
- ISSN : 0003-9861
- Web of Science ID : WOS:000182944100014