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- W4252205280 abstract "Regulation of the μM-calcium–requiring form of calpain (μ calpain) was studied in SH-SY-5Y human neuroblastoma cells. Immunoblot analysis demonstrated that the vast majority of μ calpain is localized within cytosolic pools. Calpain activation was monitored as a function of autolysis within intact cells following calcium influx from the culture medium by calcium ionophores A23187 or ionomycin, or following release of calcium from intracellular stores by thapsigargin. Within intact neuronal cells, following an influx of calcium into the cytosolic from either extracellular or intracellular sources, μ calpain is preferentially activated at the plasma membrane as evidenced by autolytic generation of faster-migrating isoforms. By contrast, similar autolytic profiles for μ calpain in membrane or cytosolic fractions following addition of calcium were observed under cell-free conditions and within cells following death due to extended ionophore-mediated calcium influx. These differential activation profiles for cytosolic μ calpain within living cells and following cellular fractionation/cell death indicate the presence of a regulatory system within neuronal cells. As in previous studies in other systems, we demonstrate the presence of a calpain activator protein. Cycloheximide treatment depleted the autolytic capacity of membrane-associated μ calpain within 4–6 hr without a corresponding decline in total μ calpain protein levels, indicating that the activator protein undergoes rapid turnover in comparison to calpain; pulse-chase radiolabeling confirmed the half-life of μ calpain to exceed 24 hr. Our data suggest that this labile protein represents a major rate-limiting step for in situ calpain activation within neuronal cells, and that, given the tremendous latent μ calpain activity within the cytosol, the interplay of the activator protein and the endogenous inhibitor calpastatin are crucial for maintaining neuronal homeostasis. J. Neurosci. Res. 48:543–550, 1997. © 1997 Wiley-Liss Inc." @default.
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- W4252205280 date "1997-06-15" @default.
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- W4252205280 title "Restriction of μM‐calcium–requiring calpain activation to the plasma membrane in human neuroblastoma cells: Evidence for regionalized influence of a calpain activator protein" @default.
- W4252205280 doi "https://doi.org/10.1002/(sici)1097-4547(19970615)48:6<543::aid-jnr7>3.3.co;2-z" @default.
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