Expression of human inducible nitric oxide synthase in Escherichia coli.
We have expressed active full-length human inducible nitric oxide synthase (iNOS) in E. coli. Expression required co-expression with calmodulin, a particularly tight-binding cofactor. The extracts also required tetrahydrobiopterin to display activity. Specific activity of the purified recombinant iNOS was similar to iNOS purified from murine macrophages. This result indicates that no special processing events unique to eucaryotic cells are necessary for iNOS activity.
Fossetta
J D
JD
Department of Immunology, Schering-Plough Research Institute, Kenilworth, NJ 07003, USA.
Niu
X D
XD
Lunn
C A
CA
Zavodny
P J
PJ
Narula
S K
SK
Lundell
D
D
D001483:Base Sequence; D001708:Biopterins / Q000031:analogs & derivatives / Q000494:pharmacology; D001921:Brain / Q000201:enzymology*; D002147:Calmodulin / Q000096:biosynthesis / Q000502:physiology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism
D002147:Calmodulin; D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D011994:Recombinant Proteins; D001708:Biopterins; D019001:Nitric Oxide Synthase; C003402:sapropterin