In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD

Di Bartolo, Natalie D., Hvorup, Rikki N., Locher, Kaspar P. and Booth, Paula J. (2011) In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD. Journal of Biological Chemistry, 286 21: 18807-18815. doi:10.1074/jbc.M110.176891

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Author Di Bartolo, Natalie D.
Hvorup, Rikki N.
Locher, Kaspar P.
Booth, Paula J.
Title In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD
Formatted title
In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
1083-351X
Publication date 2011-05-27
Sub-type Article (original research)
DOI 10.1074/jbc.M110.176891
Open Access Status File (Publisher version)
Volume 286
Issue 21
Start page 18807
End page 18815
Total pages 9
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Subject 1303 Specialist Studies in Education
1307 Cell Biology
1312 Molecular Biology
Formatted abstract
Studies on membrane protein folding have focused on monomeric α-helical proteins and a major challenge is to extend this work to larger oligomeric membrane proteins. Here, we study the Escherichia coli (E. coli) ATP-binding cassette (ABC) transporter that imports vitamin B12 (the BtuCD protein) and use it as a model system for investigating the folding and assembly of a tetrameric membrane protein complex. Our work takes advantage of the modular organization of BtuCD, which consists of two transmembrane protein subunits, BtuC, and two cytoplasmically located nucleotide-binding protein subunits, BtuD. We show that the BtuCD transporter can be re-assembled from both prefolded and partly unfolded, urea denatured BtuC and BtuD subunits. The in vitro re-assembly leads to a BtuCD complex with the correct, native, BtuC and BtuD subunit stoichiometry. The highest rates of ATP hydrolysis were achieved for BtuCD re-assembled from partly unfolded subunits. This supports the idea of cooperative folding and assembly of the constituent protein subunits of the BtuCD transporter. BtuCD folding also provides an opportunity to investigate how a protein that contains both membrane-bound and aqueous subunits coordinates the folding requirements of the hydrophobic and hydrophilic subunits.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Chemistry and Molecular Biosciences
 
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