Expression of biologically active human calpastatin in baculovirus-infected insect cells and in Escherichia coli.
Calpastatin, an endogeneous inhibitor protein acting on calpain (Ca(2+)-dependent cysteine proteinase), is widely distributed in animal tissues and cells. Two different expression systems, baculovirus-infected Spodoptera frugiperda (Sf9) insect cells and Escherichia coli, were used for overexpression of the human calpastatin tagged with N-terminal hexahistidine peptide. Recombinant calpastatin was purified to homogeneity by nickel ion affinity chromatography and gel filtration separation. Purified recombinant proteins from both systems have similar inhibitory activity for calpain.
Hitomi
K
K
School of Agricultural Sciences, Nagoya University, Japan.
Yokoyama
A
A
Maki
M
M
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors*; D002460:Cell Line; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D006639:Histidine; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D018411:Spodoptera / Q000166:cytology
D002135:Calcium-Binding Proteins; D015853:Cysteine Proteinase Inhibitors; D011993:Recombinant Fusion Proteins; D006639:Histidine; C033668:calpastatin; D002154:Calpain