Cloning and expression of an interleukin-1 beta precursor and its conversion to interleukin-1 beta.
A gene coding for a N-terminal precursor of interleukin-1 beta (IL-1 beta) was cloned and expressed in E. coli. The isolated Met-Glu-Ala-Glu-IL-1 beta precursor was enzymatically converted to IL-1 beta by means of dipeptidylaminopeptidase (DAP I). This method ensured a correct N-terminal residue and the often observed expression of Met-IL-1 beta was thus avoided. The pure and physically homogeneous product exhibited the characteristic properties of natural IL-1 beta. The in vitro biological activity was measured in the lymphocyte-activating factor assay and was compared to that of natural IL-1 beta isolated from stimulated monocyte culture using exactly the same purification procedure. The specific biological activity of both products was 2 x 10(-8) U/mg indicating that the recombinant product exhibits full biological activity.
Dalbøge
H
H
Department of Biotechnology, Nordisk Gentofte, Denmark.
Bayne
S
S
Christensen
T
T
Hejnaes
K R
KR
D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D003001:Cloning, Molecular; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D007375:Interleukin-1 / Q000235:genetics* / Q000378:metabolism; D008213:Lymphocyte Activation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
D000596:Amino Acids; D007375:Interleukin-1; D011498:Protein Precursors; D011994:Recombinant Proteins; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases