Coding
BFR

Part:BBa_K1438000

Designed by: Johann Bauerfeind   Group: iGEM14_Berlin   (2014-09-13)
Revision as of 08:02, 17 October 2014 by JohannB (Talk | contribs) (Usage and Biology)

Bacterioferritin (BFR)

Bacterioferritins are the E. coli cells natural iron storage proteins. These hollow nearly spherical protein shells detoxify the cell by sequestering excessive iron and forming Iron(III)hydroxid-oxide particels.

Bacterioferritin is an heam containing bacterial ferritin. Each heme is bound in a pocked formed by the interface between a pair of symmetry-related subunits [1]. However, it was investigated that these heme groups may be involved in the release of iron out of the ferritin iron core by forming an heme-mediated electron transfer to reduce immobilized Fe3+ to more soluble Fe2+.

[1] Frolow F, Kalb AJ, Yariv J. Structure of a unique twofold symmetric haem-binding site. Nat Struct Biol. 1994 Jul;1(7):453-60. PubMed PMID: 7664064.

[2] Yao H, Wang Y, Lovell S, Kumar R, Ruvinsky AM, Battaile KP, Vakser IA, Rivera M. The structure of the BfrB-Bfd complex reveals protein-protein interactions enabling iron release from bacterioferritin. J Am Chem Soc. 2012 Aug 15;134(32):13470-81. doi: 10.1021/ja305180n. Epub 2012 Aug 1. PubMed PMID: 22812654; PubMed Central PMCID: PMC3428730.



Usage and Biology

SDS PAGE

We succesfully expressed bacterioferritin in E. coli DH10b using our BFR expression Device ( BBa_K1438020 )

After induction with 1mM IPTG bacterioferritin (BFR) was expressed in DH10b over night. Bacterioferritin subunits were expected at a molecular weight of 19,7 kDa


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Improving BFRs Iron Storage Properties

We constructed a improved version of this protein by site-directed Mutagenesis (M52H) and add it to the parts registry as BBa_K1438001.

== Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 34
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 142


[edit]
Categories
//binding/metal
//cds
//function/biosynthesis/heme
Parameters
protein19,7 kDa