Soluble, tag-less form of the human transferrin receptor 1 ectodomain (TfR1), residues 121-760.
Transferrin, also known as (Cluster of Differentiation 71) (CD71), or p90, sTfR, is a protein that in humans is encoded by the TFRC gene.Transferrins are bilobal iron-binding glycoproteins that are responsible for tightly binding two atoms of iron acquired from the diet for transport through blood and delivery to cells. Iron-saturated transferrin binds to transferrin receptors on the cell surface, and the complex is internalized by receptor-mediated endocytosis. The transferrin receptor system is one of the most well characterized pathways for cell uptake and recombinant forms of transferrin have been used to deliver short peptides and toxins into cells. Furthermore the transferrin receptor is targeted by a number of "New World" viruses during early steps of infection (eg. Machupo virus).
From the laboratory of Jonathan Abraham, MD, PhD, Harvard University.
|Name:||Soluble human transferrin receptor (sTFR) 1|
|Accession ID:||GenBank: NM_003234.3|
|Source:||Expressed as a secreted protein HEK 293T cells. Purified using protocol described in Abraham J et al. Nat Struc Mol Biol 2010 and Lawrence CM et al Science 1999.|
|Molecular Weight:||Calculated mass of sTfR1 (residues 121-760) is 71,726 Da.|
|Amino Acid Sequence:||RLYWDDLKRKLSEKLDSTDFTSTIKLLNENSYVPREAGSQKDENLALYVENQFREFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGTGDPYTPGFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVSNVLKEIKILNIFGVIKGFVEPDHYVVVGAQRDAWGPGAAKSGVGTALLLKLAQMFSDMVLKDGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYINLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQFLYQDSNWASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMDTYKELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERYNSQLLSFVRDLNQYRADIKEMGLSLQWLYSARGDFFRATSRLTTDFGNAEKTDRFVMKKLNDRVMRVEYHFLSPYVSPKESPFRHVFWGSGSHTLPALLENLKLRKQNNGAFNETLFRNQLALATWTIQGAANALSGDVWDIDNEF|
|Buffer:||25 mM tris hydrochloride (pH 7.5), 150 mM sodium chloride|
|Storage:||+4C. Do not freeze|
We designed a construct encoding a soluble, tag-less form of the human transferrin receptor 1 ectodomain (TfR1, GenBank: NM_003234.3) by cloning residues 121-760 of TfR1 (sTfR1) into the pHL-sec expression vector1 downstream of the pHL-sec secretion signal (Fig. 1A below). We produced sTfR1 in HEK 293S GnTI-/- cells (ATCC CRL-3022) grown in suspension culture and maintained in FreeStyle 293 Expression Medium (Gibco) supplemented with 2 % Ultralow IgG FBS (ThermoFisher) and penicillin/streptomycin. For large-scale protein production, we transiently transfected HEK 293S GnTI-/- cells with the sTfR1-expressor construct using polyethylenimine. We harvested supernatant 72 hours post-transfection and purified TfR1 from supernatant using human transferrin affinity chromatography as previously described2,3. We eluted the protein with buffer containing 2M potassium chloride, 50 mM HEPES pH 7.5 (Fig. 1B below). Purified sTfR1 elutes a single peak by size exclusion chromatography (SEC) on a Superdex 200 column (GE Healthcare) (Fig. 1C below).
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