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PDBsum entry 1b3e
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Iron transport
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PDB id
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1b3e
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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X-Ray crystallography and mass spectroscopy reveal that the n-Lobe of human transferrin expressed in pichia pastoris is folded correctly but is glycosylated on serine-32.
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Authors
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M.C.Bewley,
B.M.Tam,
J.Grewal,
S.He,
S.Shewry,
M.E.Murphy,
A.B.Mason,
R.C.Woodworth,
E.N.Baker,
R.T.Macgillivray.
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Ref.
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Biochemistry, 1999,
38,
2535-2541.
[DOI no: ]
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PubMed id
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Abstract
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The ferric form of the N-lobe of human serum transferrin (Fe(III)-hTF/2N) has
been expressed at high levels in Pichia pastoris. The Fe(III)-hTF/2N was
crystallized in the space group P41212, and X-ray crystallography was used to
solve the structure of the recombinant protein at 2.5 A resolution. This
represents only the second P. pastoris-derived protein structure determined to
date, and allows the comparison of the structures of recombinant Fe(III)-hTF/2N
expressed in P. pastoris and mammalian cells with serum-derived transferrin. The
polypeptide folding pattern is essentially identical in all of the three
proteins. Mass spectroscopic analyses of P. pastoris- hTF/2N and proteolytically
derived fragments revealed glycosylation of Ser-32 with a single hexose. This
represents the first localization of an O-linked glycan in a P. pastoris-derived
protein. Because of its distance from the iron-binding site, glycosylation of
Ser-32 should not affect the iron-binding properties of hTF/2N expressed in P.
pastoris, making this an excellent expression system for the production of
hTF/2N.
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