Expression and purification of human interleukin-1beta converting enzyme from Trichoplusia ni insect cells using a baculovirus expression system.
Employing a baculovirus expression system, human interleukin-1beta converting enzyme (ICE) has been expressed in Trichoplusia ni High-Five insect cells and purified. ICE was expressed with an N-terminal T7 epitope, thus allowing its purification using immobilized anti-T7 antibodies. The recombinant ICE was purified to >95% homogeneity, the one minor contaminant being the baculovirus anti-apoptotic protein (P35) which was copurified. The purified recombinant ICE was biologically active, cleaving both pIL-1beta and poly(ADP-ribose) polymerase substrates. The kinetic properties of the purified recombinant ICE compare favorably with native ICE purified from a THP-1 monocytic line.
Chen
W
W
Vertex Pharmaceuticals, Inc., Cambridge, Massachusetts 02139-4242, USA.
Raybuck
S A
SA
Fulghum
J R
JR
Petrillo
R A
RA
Margolin
N
N
Chambers
S P
SP
D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics / Q000276:immunology; D020170:Caspase 1; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000939:Epitopes; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D007700:Kinetics; D009036:Moths / Q000166:cytology / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D017421:Sequence Analysis; D014764:Viral Proteins / Q000302:isolation & purification
D000939:Epitopes; D051034:Inhibitor of Apoptosis Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C086430:inhibitor of apoptosis, Nucleopolyhedrovirus; D003546:Cysteine Endopeptidases; D020170:Caspase 1