PMID- 10029548
OWN - NLM
STAT- MEDLINE
DCOM- 19990316
LR  - 20131121
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 8
DP  - 1999 Feb 23
TI  - 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.
PG  - 2535-41
AB  - 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.
FAU - Bewley, M C
AU  - Bewley MC
AD  - Institute of Molecular Biosciences, College of Sciences, Massey University,
      Palmerston North, New Zealand.
FAU - Tam, B M
AU  - Tam BM
FAU - Grewal, J
AU  - Grewal J
FAU - He, S
AU  - He S
FAU - Shewry, S
AU  - Shewry S
FAU - Murphy, M E
AU  - Murphy ME
FAU - Mason, A B
AU  - Mason AB
FAU - Woodworth, R C
AU  - Woodworth RC
FAU - Baker, E N
AU  - Baker EN
FAU - MacGillivray, R T
AU  - MacGillivray RT
LA  - eng
SI  - PDB/1B3E
GR  - R01 DK 21739/DK/NIDDK NIH HHS/United States
GR  - R01 DK 35533/DK/NIDDK NIH HHS/United States
GR  - R01 HD 20859/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Ferric Compounds)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Transferrin)
RN  - 452VLY9402 (Serine)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cricetinae
MH  - Crystallization
MH  - Crystallography, X-Ray
MH  - Ferric Compounds/chemistry
MH  - Glycosylation
MH  - Humans
MH  - Kidney/cytology
MH  - Mass Spectrometry
MH  - Models, Molecular
MH  - Peptide Fragments/*chemistry/genetics/metabolism
MH  - Pichia/*genetics
MH  - *Protein Folding
MH  - Recombinant Proteins/biosynthesis/*chemistry/metabolism
MH  - Serine/genetics/*metabolism
MH  - Transferrin/*chemistry/genetics/metabolism
EDAT- 1999/02/25 00:00
MHDA- 1999/02/25 00:01
CRDT- 1999/02/25 00:00
PHST- 1999/02/25 00:00 [pubmed]
PHST- 1999/02/25 00:01 [medline]
PHST- 1999/02/25 00:00 [entrez]
AID - 10.1021/bi9824543 [doi]
AID - bi9824543 [pii]
PST - ppublish
SO  - Biochemistry. 1999 Feb 23;38(8):2535-41. doi: 10.1021/bi9824543.