Non-erythroid alpha-spectrin breakdown by calpain and interleukin 1 beta-converting-enzyme-like protease(s) in apoptotic cells: contributory roles of both protease families in neuronal apoptosis.
The cytoskeletal protein non-erythroid alpha-spectrin is well documented as an endogenous calpain substrate, especially under pathophysiological conditions. In cell necrosis (e.g. maitotoxin-treated neuroblastoma SH-SY5Y cells), alpha-spectrin breakdown products (SBDPs) of 150 kDa and 145 kDa were produced by cellular calpains. In contrast, in neuronal cells undergoing apoptosis (cerebellar granule neurons subjected to low potassium and SH-SY5Y cells treated with staurosporine), an additional SBDP of 120 kDa was also observed. The formation of the 120 kDa SBDP was insensitive to calpain inhibitors but was completely blocked by an interleukin 1 beta-converting-enzyme (ICE)-like protease inhibitor, Z-Asp-CH2OC(O)-2,6-dichlorobenzene. Autolytic activation of both calpain and the ICE homologue CPP32 was also observed in apoptotic cells. alpha-Spectrin can also be cleaved in vitro by purified calpains to produce the SBDP doublet of 150/145 kDa and by ICE and ICE homologues [ICH-1, ICH-2 and CPP32(beta)] to produce a 150 kDa SBDP. In addition, CPP32 and ICE also produced a 120 kDa SBDP. Furthermore inhibition of either ICE-like protease(s) or calpain protects both granule neurons and SH-SY5Y cells against apoptosis. Our results suggest that both protease families participate in the expression of neuronal apoptosis.
Nath
R
R
Department of Neuroscience Therapeutics, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI 48105, USA.
Raser
K J
KJ
Stafford
D
D
Hajimohammadreza
I
I
Posner
A
A
Allen
H
H
Talanian
R V
RV
Yuen
P
P
Gilbertsen
R B
RB
Wang
K K
KK
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D002154:Calpain / Q000378:metabolism*; D020170:Caspase 1; D002460:Cell Line; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000201:enzymology; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008387:Marine Toxins / Q000494:pharmacology; D008969:Molecular Sequence Data; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D035964:Oxocins; D016716:PC12 Cells; D051381:Rats; D013049:Spectrin / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured; D000095702:Polyether Toxins
D002154:Calpain; D020170:Caspase 1; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors; D008387:Marine Toxins; D035964:Oxocins; D013049:Spectrin; D019311:Staurosporine; C020394:maitotoxin; D000095702:Polyether Toxins