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 tran...

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Bibliographic Details
Published in:The international journal of biochemistry & cell biology 1995-08, Vol.27 (8), p.839-850
Main Authors: de Smit, Maarten H., Hoefkens, Peter, de Jong, Gerard, van Duin, Jan, van Knippenberg, Peter H., van Eijk, Hendrik G.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Binding Sites
Escherichia coli
Gene expression
Genetic Vectors
Glycosylation
Humans
Inclusion bodie
Molecular Sequence Data
Peptide Fragments - biosynthesis
Peptide Fragments - genetics
Plasmids
Protein degradation
Recombinant Proteins - biosynthesis
Ribosomes - metabolism
Transferrin - biosynthesis
Transferrin - genetics
Transferrin-cDNA sequence
Translational coupling
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Summary: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/1. 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.
ISSN:1357-2725
1878-5875
DOI:10.1016/1357-2725(95)00040-V