Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae

Peter J. Sargent; Sebastien Farnaud; Richard Cammack; Heinz M P Zoller; Robert W. Evans

doi:10.1007/s10534-005-5532-6

Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae

Peter J. Sargent, Sebastien Farnaud, Richard Cammack, Heinz M P Zoller, Robert W. Evans

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

Structural identity between a recombinant transferrin mutant (N413Q, N611Q) secreted from Saccharomyces cerevisiae and the native protein was shown by CD analysis and immunodiffusion assays against anti-hSTf. The ability of the recombinant protein to bind iron was confirmed by urea-PAGE and EPR analysis of the iron-saturated protein revealed the characteristic holo-transferrin spectrum, indicating conservation of both iron-binding sites. The integrity of the unglycosylated recombinant protein indicates that such protein could be a valuable tool not only for structure-function characterisation but also crystallisation assays. In addition, the recombinant transferrin was found to be as effective as native transferrin as a growth factor in cell culture medium.

Original language	English
Pages (from-to)	513-519
Number of pages	7
Journal	BioMetals
Volume	19
Issue number	5
DOIs	https://doi.org/10.1007/s10534-005-5532-6
Publication status	Published - Oct 2006
Externally published	Yes

Keywords

Characterisation
Iron
Recombinant
Saccharomyces cerevisiae
Transferrin
Yeast

ASJC Scopus subject areas

General Agricultural and Biological Sciences
General Biochemistry,Genetics and Molecular Biology

Access to Document

10.1007/s10534-005-5532-6

Cite this

@article{e9ef25b6a67249a3b6d5e10ded29976c,

title = "Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae",

abstract = "Structural identity between a recombinant transferrin mutant (N413Q, N611Q) secreted from Saccharomyces cerevisiae and the native protein was shown by CD analysis and immunodiffusion assays against anti-hSTf. The ability of the recombinant protein to bind iron was confirmed by urea-PAGE and EPR analysis of the iron-saturated protein revealed the characteristic holo-transferrin spectrum, indicating conservation of both iron-binding sites. The integrity of the unglycosylated recombinant protein indicates that such protein could be a valuable tool not only for structure-function characterisation but also crystallisation assays. In addition, the recombinant transferrin was found to be as effective as native transferrin as a growth factor in cell culture medium.",

keywords = "Characterisation, Iron, Recombinant, Saccharomyces cerevisiae, Transferrin, Yeast",

author = "Sargent, {Peter J.} and Sebastien Farnaud and Richard Cammack and Zoller, {Heinz M P} and Evans, {Robert W.}",

year = "2006",

month = oct,

doi = "10.1007/s10534-005-5532-6",

language = "English",

volume = "19",

pages = "513--519",

journal = "BioMetals",

issn = "0966-0844",

publisher = "Springer Verlag",

number = "5",

}

TY - JOUR

T1 - Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae

AU - Sargent, Peter J.

AU - Farnaud, Sebastien

AU - Cammack, Richard

AU - Zoller, Heinz M P

AU - Evans, Robert W.

PY - 2006/10

Y1 - 2006/10

N2 - Structural identity between a recombinant transferrin mutant (N413Q, N611Q) secreted from Saccharomyces cerevisiae and the native protein was shown by CD analysis and immunodiffusion assays against anti-hSTf. The ability of the recombinant protein to bind iron was confirmed by urea-PAGE and EPR analysis of the iron-saturated protein revealed the characteristic holo-transferrin spectrum, indicating conservation of both iron-binding sites. The integrity of the unglycosylated recombinant protein indicates that such protein could be a valuable tool not only for structure-function characterisation but also crystallisation assays. In addition, the recombinant transferrin was found to be as effective as native transferrin as a growth factor in cell culture medium.

AB - Structural identity between a recombinant transferrin mutant (N413Q, N611Q) secreted from Saccharomyces cerevisiae and the native protein was shown by CD analysis and immunodiffusion assays against anti-hSTf. The ability of the recombinant protein to bind iron was confirmed by urea-PAGE and EPR analysis of the iron-saturated protein revealed the characteristic holo-transferrin spectrum, indicating conservation of both iron-binding sites. The integrity of the unglycosylated recombinant protein indicates that such protein could be a valuable tool not only for structure-function characterisation but also crystallisation assays. In addition, the recombinant transferrin was found to be as effective as native transferrin as a growth factor in cell culture medium.

KW - Characterisation

KW - Iron

KW - Recombinant

KW - Saccharomyces cerevisiae

KW - Transferrin

KW - Yeast

UR - http://www.scopus.com/inward/record.url?scp=33747894122&partnerID=8YFLogxK

U2 - 10.1007/s10534-005-5532-6

DO - 10.1007/s10534-005-5532-6

M3 - Article

C2 - 16937257

AN - SCOPUS:33747894122

SN - 0966-0844

VL - 19

SP - 513

EP - 519

JO - BioMetals

JF - BioMetals

IS - 5

ER -

Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this