Expression of human parathyroid hormone in Escherichia coli.
Human parathyroid hormone (PTH) has been expressed in Escherichia coli as a cro-beta-galactosidase-hPTH fusion protein under temperature-sensitive control of the lambda phage PR promoter. The lacZ gene has been truncated to a different extent revealing an optimal length of the prokaryotic peptide portion between 199 and 407 amino acid residues. Up to 250 mg of pure fusion protein have been obtained from 1-liter E. coli culture by stepwise solubilization with urea. The linkage between the prokaryotic and the eukaryotic protein moiety consists of an Asp-Pro peptide bond and therefore is easily cleavable by acid treatment. A simple procedure for the purification of the hormone is described. The resulting recombinant hormone reacts with anti-PTH antibodies and stimulates renal adenylate cyclase identically to bovine or human PTH.
Wingender
E
E
Gesellschaft für Biotechnologische Forschung, Braunschweig, Federal Republic of Germany.
Bercz
G
G
Blöcker
H
H
Frank
R
R
Mayer
H
H
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005561:Formates; D005822:Genetic Vectors; D006801:Humans; D006868:Hydrolysis; D007668:Kidney / Q000187:drug effects / Q000201:enzymology; D010281:Parathyroid Hormone / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics*; D013552:Swine; D013696:Temperature; D001616:beta-Galactosidase / Q000235:genetics
D005561:Formates; D010281:Parathyroid Hormone; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C030544:formic acid; D001616:beta-Galactosidase; D000262:Adenylyl Cyclases