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J Biol Chem, Vol. 273, Issue 35, 22490-22497, August 28, 1998
From the ¶ Laboratory of Neuro-biochemistry, The degradation of
Simultaneous Degradation of
II- and 
II-Spectrin by Caspase
3 (CPP32) in Apoptotic Cells
¶,
,,, Immunopathology
and ** Chemistry, Parke-Davis Pharmaceutical Research, Division of
Warner-Lambert Company, Ann Arbor, Michigan 48105, the
Department of Pharmacology, University of Michigan
Medical School, Ann Arbor, Michigan 48106, the
BASF Bioresearch Corporation, Worcester,
Massachusetts 01605, and the §§ Department of
Pathology, Yale University School of Medicine,
New Haven, Connecticut 06510
II- and 
II-spectrin
during apoptosis in cultured human neuroblastoma SH-SY5Y cells was
investigated. Immunofluorescent staining showed that the collapse of
the cortical spectrin cytoskeleton is an early event following
staurosporine challenge. This collapse correlated with the generation
of a series of prominent spectrin breakdown products (BDPs) derived
from both II- and II-subunits. Major C-terminal II-spectrin
BDPs were detected at 
150, 145, and 120 kDa (II-BDP150,
II-BDP145, and II-BDP120, respectively); major C-terminal
II-spectrin BDPs were at 110 and 85 kDa (II-BDP110 and
II-BDP85, respectively). N-terminal sequencing of the major
fragments produced in vitro by caspase 3 revealed that
II-BDP150 and II-BDP120 were generated by cleavages at
DETD1185*S1186 and
DSLD1478*S1479, respectively. For
II-spectrin, a major caspase site was detected at
DEVD1457*S1458, and both
II-BDP110 and II-BDP85 shared a common N-terminal sequence
starting with Ser1458. An additional cleavage site near the
C terminus, at ETVD2146*S2147, was found to
account for II-BDP85. Studies using specific caspase or calpain
inhibitors indicate that the pattern of spectrin breakdown during
apoptosis differs from that during non-apoptotic cell death. We
postulate that in concert with calpain, caspase rapidly targets critical sites in both II- and II-spectrin and thereby initiates a rapid dissolution of the spectrin-actin cortical cytoskeleton with
apoptosis.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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