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Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae

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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.

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Authors and Affiliations

  1. Metalloprotein Research Group, Randall Division of Cell and Molecular Biophysics, King’s College London, Guy’s Campus, St Thomas Street, 3.6b New Hunts House, London, SE1 1UL, UK

    Peter J. Sargent, Sebastien Farnaud & Robert W. Evans

  2. Pharmaceutical Sciences Research Division, King’s College London, Franklin-Wilkins Building, 150 Stamford St, London, SE1 9NH, UK

    Richard Cammack

  3. Department of Medicine, University of Cambridge, Addenbrooke’s Hospital, P.O. Box 157, Level 5, Hills Road, Cambridge, CB2 2QQ, UK

    Heinz M. P. Zoller

Authors
  1. Peter J. Sargent
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  2. Sebastien Farnaud
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  3. Richard Cammack
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  4. Heinz M. P. Zoller
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  5. Robert W. Evans
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Correspondence to Peter J. Sargent.

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Sargent, P.J., Farnaud, S., Cammack, R. et al. Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae . Biometals 19, 513–519 (2006). https://doi.org/10.1007/s10534-005-5532-6

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  • DOI: https://doi.org/10.1007/s10534-005-5532-6

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