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Calcium-dependent Plasma Membrane Binding and Cell Lysis by Perforin Are Mediated through Its C2 Domain
The lymphocyte pore-forming protein perforin is essential for maintaining immune homeostasis and for effective defense against intracellular pathogens. To date, there have been no reported structure-function studies to substantiate the function of any putative domains of perforin, which have been po...
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Published in: | The Journal of biological chemistry 2005-03, Vol.280 (9), p.8426-8434 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | The lymphocyte pore-forming protein perforin is essential for maintaining immune homeostasis and for effective defense against
intracellular pathogens. To date, there have been no reported structure-function studies to substantiate the function of any
putative domains of perforin, which have been postulated totally on primary sequence similarities with domains in other proteins.
In this report, we have used recently developed modalities for expressing full-length perforin and robust functional assays
to investigate one of the hallmarks of perforin function: its absolute dependence on calcium for lipid binding and cell lysis.
We provide, for the first time, experimental evidence that the predicted C-terminal C2 motif constitutes a functional domain
that is responsible for membrane binding of perforin. Whereas conserved aspartate residues at positions 429, 435, 483, and
485 were essential for calcium-dependent plasma membrane binding and cell lysis, the contribution of Asp-491 was limited.
Finally, after experimentally verifying an optimized three-dimensional model, we have made predictions on the impact of two
inherited perforin mutations of the C2 domain on calcium-dependent lipid binding and cell lysis. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M413303200 |