Characterization of a legumain/vacuolar processing enzyme and YVADase activity in Papaver pollen

Abstract

Legumains, also known as Vacuolar Processing Enzymes (VPEs) have received considerable attention recently, as they share structural properties with mammalian caspase-1 and exhibit YVADase/caspase-1-like cleavage activity. Although many legumains have been cloned, knowledge about their detailed characteristics and intracellular localization is relatively limited. We previously identified several caspase-like activities activated by self-incompatibility (SI) in pollen; a DEVDase was required for programmed cell death (PCD), but YVADase was not (Bosch and Franklin-Tong in Proc Natl Acad Sci USA 104:18327–18332, 2007; Thomas and Franklin-Tong in Nature 429:305–309, 2004). Here we report identification of a legumain/VPE from Papaver rhoeas pollen (PrVPE1) that binds to the DEVD tetrapeptide, a signature substrate for caspase-3. A detailed characterization of the recombinant PrVPE1 cleavage activity revealed that, like other VPEs, it has YVADase activity and requires an acidic pH for activity. Unlike other legumain/VPEs, it also exhibits DEVDase and IETDase activities and apparently does not require processing for activity. The pollen-expressed PrVPE1 localizes to a reticulate compartment resembling the vacuole. Examination of YVADase activity using live-cell imaging of pollen tubes revealed YVADase activity in mitochondria of growing pollen tubes. The unexpected features of PrVPE1, together with evidence for YVADase activity in plant mitochondria, indicate that VPEs, YVADases, their localization and functions in plant cells merit further investigation.

DOI: 10.1007/s11103-010-9681-9

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Cite this paper

@article{Bosch2010CharacterizationOA, title={Characterization of a legumain/vacuolar processing enzyme and YVADase activity in Papaver pollen}, author={Maurice Bosch and Natalie S Poulter and Ruth M. Perry and Katie A Wilkins and Vernonica E Franklin-Tong}, journal={Plant Molecular Biology}, year={2010}, volume={74}, pages={381-393} }