1B3E

HUMAN SERUM TRANSFERRIN, N-TERMINAL LOBE, EXPRESSED IN PICHIA PASTORIS

Summary for 1B3E

• Entry DOI: 10.2210/pdb1b3e/pdb
• Descriptor: PROTEIN (SERUM TRANSFERRIN), FE (III) ION, CARBONATE ION, ... (4 entities in total)
• Functional Keywords: iron transport, glycoprotein, transferrin, pichia pastoris, glycosylation
• Biological source: Homo sapiens (human)
• Cellular location: Secreted: P02787
• Total number of polymer chains: 1
• Total formula weight: 36621.38

Authors

Bewley, M.C.,Tam, B.M.,Grewal, J.,He, S.,Shewry, S.,Murphy, M.E.P.,Mason, A.B.,Woodworth, R.C.,Baker, E.N.,Macgillivray, R.T.A. (deposition date: 1998-12-09, release date: 1999-03-26, Last modification date: 2024-10-30)

Primary citation

Bewley, M.C.,Tam, B.M.,Grewal, J.,He, S.,Shewry, S.,Murphy, M.E.,Mason, A.B.,Woodworth, R.C.,Baker, E.N.,MacGillivray, R.T.
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.
Biochemistry, 38:2535-2541, 1999

PubMed Abstract: 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.

PubMed: 10029548
DOI: 10.1021/bi9824543

Experimental method

X-RAY DIFFRACTION (2.5 Å)