Isolation, renaturation and partial characterization of recombinant human transferrin and its half molecules from Escherichia coli.
Recombinant human transferrin as well as N- and C-terminal half-transferrins, produced in Escherichia coli, are deposited in inclusion bodies by the bacteria. The isolation and purification of the recombinant proteins from these inclusion bodies are described here. The amino acid compositions and N-terminal sequences of the proteins were determined, and found to be in agreement with the known protein structure of human serum transferrin. Renaturation of the recombinant proteins is described, resulting in water-soluble iron-binding molecules. Iron binding was confirmed by 59Fe labelling, absorption spectrophotometry and EPR spectrometry.
Hoefkens
P
P
Department of Chemical Pathology, Erasmus University, Rotterdam, The Netherlands.
de Smit
M H
MH
de Jeu-Jaspars
N M
NM
Huijskes-Heins
M I
MI
de Jong
G
G
van Eijk
H G
HG
D000595:Amino Acid Sequence; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014168:Transferrin / Q000737:chemistry / Q000302:isolation & purification*
D011994:Recombinant Proteins; D014168:Transferrin