Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
Protein Expr Purif. 2008 Apr;58(2):229-41. doi: 10.1016/j.pep.2007.11.018. Epub 2007 Dec 15.

A Protein Structure Initiative approach to expression, purification, and in situ delivery of human cytochrome b5 to membrane vesicles.

Author information

  • 1Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Room 141B, 433 Babcock Drive, Madison, WI 53706, USA.

Abstract

A specialized vector backbone from the Protein Structure Initiative was used to express full-length human cytochrome b5 as a C-terminal fusion to His8-maltose binding protein in Escherichia coli. The fusion protein could be completely cleaved by tobacco etch virus protease, and a yield of approximately 18 mg of purified full-length human cytochrome b5 per liter of culture medium was obtained (2.3mg per g of wet weight bacterial cells). In situ proteolysis of the fusion protein in the presence of chemically defined synthetic liposomes allowed facile spontaneous delivery of the functional peripheral membrane protein into a defined membrane environment without prior exposure to detergents or other lipids. The utility of this approach as a delivery method for production and incorporation of monotopic (peripheral) membrane proteins is discussed.

PMID:
18226920
[PubMed - indexed for MEDLINE]
PMCID:
PMC2277500
Free PMC Article

Images from this publication.See all images (7)Free text

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk