Characterisation of recombinant unglycosylated human serum transferrin purified from Saccharomyces cerevisiae.
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.
Sargent
Peter J
PJ
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.sargent@kcl.ac.uk
Farnaud
Sebastien
S
Cammack
Richard
R
Zoller
Heinz M P
HM
Evans
Robert W
RW
D049109:Cell Proliferation; D004578:Electron Spin Resonance Spectroscopy; D006031:Glycosylation; D006801:Humans; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013057:Spectrum Analysis; D014168:Transferrin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D014168:Transferrin