Cell-free synthesis and characterization of a novel cytotoxic pierisin-like protein from the cabbage butterfly Pieris rapae.
Pierisin-like proteins comprise a growing family of ADP-ribosyltransferases expressed in various species of white butterflies. The prototype pierisin-1 from the cabbage butterfly, Pieris rapae, was identified as a potent apoptosis-inducing agent, acting on various types of carcinoma cell lines by mono-ADP-ribosylation of DNA. The characterization of pierisin-like proteins is hampered by its potent toxicity, which prevents its expression as a recombinant protein in Escherichia coli. Here we characterized a new member of the pierisin protein family named pierisin-1b, which was cloned from P. rapae. Pierisin-1b consists of 849 amino acids residues and shares 63%-91% identity with already described pierisins. For expression of pierisin-1b a novel in vitro translation system was utilized. Obtained protein exhibits specific ADP-ribosyltransferase activity on deoxyguanosine residues of DNA leading to induction of apoptosis and cell death.
Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, Albertstr. 25, D-79104 Freiburg, Germany. Joachim.Orth@pharmakol.uni-freiburg.de
Joachim H C
JH
Orth
Björn
B
Schorch
Sam
S
Boundy
Richard
R
Ffrench-Constant
Stefan
S
Kubick
Klaus
K
Aktories
D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002080:Butterflies / Q000737:chemistry*; D002474:Cell-Free System; D003001:Cloning, Molecular; D003603:Cytotoxins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D003603:Cytotoxins; D019476:Insect Proteins; C113557:pierisin protein, insect; D036002:ADP Ribose Transferases