Actin is cleaved during constitutive apoptosis.
Proteases play an important role in the programme of cell death by apoptosis but little is known of the substrates cleaved, particularly in constitutive models of this type of cell death. Neutrophils spontaneously undergo apoptosis in culture without requiring external stimuli. During this process we found biochemical and immunochemical evidence for the cleavage of membrane-associated actin, a component of the cytoskeleton that links polymerized actin to the plasma membrane. Cleavage occurred at a single site at the N-terminus, between residues Val43-Met44, a site devoid of a consensus motif for cleavage by cysteine proteases of the interleukin-1beta-converting enzyme (ICE)-family. Whereas actin cleavage and nuclear/cell surface markers of apoptosis were co-ordinately diminished by zVAD-fmk, an inhibitor of the ICE-like family of proteases, only acetyl-leucyl-leucylnormethional, an inhibitor of calpains, was capable of completely inhibiting actin cleavage. Our results suggest that actin is not a direct substrate for the ICE-like family of proteases. By disabling the cytoskeleton, actin cleavage may be an important component in the capacity of apoptosis to reduce the injurious potential of neutrophils.
Brown
S B
SB
Division of Renal and Inflammatory Disease, Department of Medicine, University Hospital, Nottingham NG7 2UH, UK.
Bailey
K
K
Savill
J
J
D000199:Actins / Q000378:metabolism*; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000595:Amino Acid Sequence; D017209:Apoptosis; D002154:Calpain / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology
D000199:Actins; D000590:Amino Acid Chloromethyl Ketones; D007976:Leupeptins; D011480:Protease Inhibitors; C089487:acetyl-leucinyl-leucinyl-methional; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D002154:Calpain