Expression of a pokeweed antiviral protein in Escherichia coli and its characterization.
Two expression vectors were constructed to produce a putative mature alpha-pokeweed antiviral protein (alpha-PAP) in Escherichia coli with its NH2- and COOH-terminal extrapeptides excised. One was for its intracellular expression with a methionine at its NH2-terminal. The other was for its secretion using an ompA signal peptide. The former product was purified from the total soluble proteins of the transformant with a yield of 1.74 mg/liter and the latter had a yield of 5.55 mg/liter. Both products exhibited RNA N-glycosidase activity on wheat ribosomes and inhibitory activity to protein synthesis in a rabbit reticulocyte system.
Kataoka
J
J
Life Science Research Laboratory, Japan Tobacco Inc., Kanagawa.
Ago
H
H
Habuka
N
N
Furuno
M
M
Masuta
C
C
Miyano
M
M
Koiwai
A
A
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D054788:Ribosome Inactivating Proteins; D054789:Ribosome Inactivating Proteins, Type 1
D000998:Antiviral Agents; D004277:DNA, Single-Stranded; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins; C024275:pokeweed antiviral protein