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The virally encoded fungal toxin KP4 specifically blocks L-type voltage-gated calcium channels
Matthew J Gage1, Stanley G Rane, Gregory H Hockerman
1Donald Danforth Plant Science Center, St. Louis, Missouri 63132, USA.
Abstract:
KP4 is a virally encoded fungal toxin secreted by the P4 killer strain of Ustilago maydis. Previous studies demonstrated that this toxin inhibits growth of the target fungal cells by blocking calcium uptake rather than forming channels, as had been suggested previously. Unexpectedly, this toxin was also shown to inhibit voltage-gated calcium channel activity in mammalian cells. We used whole-cell patch-clamp techniques to further characterize this activity against mammalian cells. KP4 is shown to specifically block L-type calcium channels with weak voltage dependence to the block. Because KP4 activity is abrogated by calcium, KP4 probably binds competitively with calcium to the channel exterior. Finally, it is shown that chemical reagents that modify lysine residues reduce KP4 activity in both patch-clamp experiments on mammalian cells and in fungal killing assays. Because the only lysine residue is K42, this residue seems to be crucial for both mammalian and fungal channel activity. Our results defining the type of mammalian channel affected by this fungal toxin further support our contention that KP4 inhibits fungal growth by blocking transmembrane calcium flux through fungal calcium channels, and imply a high degree of structural homology between these fungal and mammalian calcium channels.
Insights
The fungal toxin KP4 blocks calcium uptake in fungal cells and inhibits L-type calcium channels in mammalian cells. This suggests structural similarities between fungal and mammalian calcium channels, with lysine 42 being crucial for activity.
Area of Science:
- Mycology
- Molecular Biology
- Biochemistry
Background:
- KP4 is a fungal toxin from Ustilago maydis.
- It inhibits fungal growth by blocking calcium uptake.
- KP4 also affects mammalian cell calcium channels.
Purpose of the Study:
- Characterize KP4's activity on mammalian calcium channels.
- Investigate the mechanism of KP4's inhibitory action.
- Identify key residues involved in KP4's function.
Main Methods:
- Whole-cell patch-clamp electrophysiology on mammalian cells.
- Fungal growth inhibition assays.
- Chemical modification of lysine residues.
Main Results:
- KP4 specifically blocks L-type calcium channels with weak voltage dependence.
- KP4 likely binds competitively with calcium to the channel exterior.
- Modification of lysine residue K42 abrogates KP4 activity in both fungal and mammalian systems.
Conclusions:
- KP4 inhibits fungal growth by blocking calcium channels, similar to its action on mammalian L-type channels.
- There is a high degree of structural homology between fungal and mammalian calcium channels.
- Lysine 42 is critical for KP4's interaction with calcium channels in both fungi and mammals.