Expression in Escherichia coli and in vitro refolding of the human protein pLG72.
Recently, genes coding for pLG72 and d-amino acid oxidase have been related to schizophrenia, a widespread psychiatric disorder that affects about 1% of population. pLG72 is a puzzling, novel protein present only in primates and proposed to be an activator of d-amino acid oxidase. Here we report on the overexpression of wild-type and His-tagged pLG72 in Escherichia coli. Both variants form inclusion bodies and have been refolded and purified to homogeneity: the acquisition of secondary and tertiary structure was demonstrated by CD spectroscopy. A figure of approximately 70 mg of pure protein per liter of fermentation broth was achieved.
Molla
Gianluca
G
Department of Biotechnology and Molecular Sciences, University of Insubria, Via J. H. Dunant, 3, 21100 Varese, Italy. gianluca.molla@uninsubria.it
Bernasconi
Mariagrazia
M
Sacchi
Silvia
S
Pilone
Mirella S
MS
Pollegioni
Loredano
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003605:D-Amino-Acid Oxidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D008969:Molecular Sequence Data; D017510:Protein Folding; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D002352:Carrier Proteins; C475071:DAOA protein, human; D017931:DNA Primers; D047908:Intracellular Signaling Peptides and Proteins; D011994:Recombinant Proteins; D003605:D-Amino-Acid Oxidase