1. Anchoring proteins to Escherichia coli cell membranes using hydrophobic anchors derived from a Bacillus subtilis integral membrane protein.
- Author
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Yang C, Xie H, Zhang JK, and Su BL
- Subjects
- Alkaline Phosphatase genetics, Alkaline Phosphatase metabolism, Amino Acid Sequence, Bacillus subtilis genetics, Bacillus subtilis metabolism, Bacterial Proteins genetics, Bacterial Proteins metabolism, Cloning, Molecular, Escherichia coli cytology, Escherichia coli genetics, Escherichia coli metabolism, Gene Expression, Genetic Vectors genetics, Hydrophobic and Hydrophilic Interactions, Membrane Proteins genetics, Membrane Proteins metabolism, Molecular Sequence Data, Periplasm chemistry, Plasmids genetics, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Alkaline Phosphatase chemistry, Bacillus subtilis chemistry, Bacterial Proteins chemistry, Escherichia coli chemistry, Membrane Proteins chemistry, Recombinant Fusion Proteins chemistry
- Abstract
Anchored periplasmic expression (APEx) technology aims to express and localize proteins or peptides in the Escherichia coli periplasm. Some reports have suggested that transmembrane segments of integral membrane proteins can be used as membrane anchors in the APEx system. In this study, a series of hydrophobic anchors derived from the first putative transmembrane helix of a Bacillus subtilis integral membrane protein, MrpF, and its truncated forms were investigated for anchored periplasmic expression of alkaline phosphatase (PhoA) in E. coli. Anchoring efficiency of hydrophobic anchors was evaluated by monitoring the expression and activity of anchored PhoA. The length of hydrophobic anchors was found to be critical for anchoring proteins to cell membranes. This study may open new avenues for applying transmembrane segments derived from native membrane proteins as membrane anchors in the APEx system., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
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