Optimized bacterial production of nonglycosylated human transferrin and its half-molecules.
Transferrin is a glycoprotein functioning in iron transport in higher eukaryotes, and consists of two highly homologous domains. To study the function of the glycan residues attached exclusively to the C-terminal domain, we have constructed a plasmid allowing production of nonglycosylated human transferrin in Escherichia coli. By molecular biological and genetic techniques, production was stepped up to 60 mg/l. Similar plasmids were constructed for production of the two half-transferrins. The recombinant proteins accumulate in inclusion-body-like aggregates, where they appear to bind iron without causing bacteriostasis. Proteins active in iron binding have been purified from these inclusion bodies.
de Smit
M H
MH
Leiden Institute of Chemistry, Department of Biochemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
Hoefkens
P
P
de Jong
G
G
van Duin
J
J
van Knippenberg
P H
PH
van Eijk
H G
HG
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004926:Escherichia coli; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012270:Ribosomes / Q000378:metabolism; D014168:Transferrin / Q000096:biosynthesis / Q000235:genetics*
D010446:Peptide Fragments; D011994:Recombinant Proteins; D014168:Transferrin