Advances in understanding pectin methylesterase inhibitor in kiwi fruit: an immunological approach
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In order to gain insight into the in situ properties and localisation of kiwi pectin methylesterase inhibitor (PMEI), a toolbox of monoclonal antibodies (MA) towards PMEI was developed. Out of a panel of MA generated towards kiwi PMEI, three MA, i.e. MA-KI9A8, MA-KI15C12 and MA-KI15G7, were selected. Thorough characterisation proved that these MA bind specifically to kiwi PMEI and kiwi PMEI in complex with plant PME and recognise a linear epitope on PMEI. Extract screening of green kiwi (Actinidia deliciosa) and gold kiwi (Actinidia chinensis) confirmed the potential use of these MA as probes to screen for PMEI in other sources. Tissue printing revealed the overall presence of PMEI in pericarp and columella of ripe kiwi fruit. Further analysis on the cellular level showed PMEI label concentrated in the middle lamella and in the cell-wall region near the plasmalemma. Intercellular spaces, however, were either completely filled or lined with label. In conclusion, the developed toolbox of antibodies towards PMEI can be used as probes to localise PMEI on different levels, which can be of relevance for plant physiologists as well as food technologists.
KeywordsImmunolocalisation Kiwi fruit Monoclonal antibodies Pectin methylesterase inhibitor
Aspergillus aculeatus pectin methylesterase
Pectin methylesterase inhibitor
Tomato (Solanum Lycopersicon) pectin methylesterase
This research has been carried out with financial support from the Research Council K.U.Leuven (IDO/05/011). S. Christiaens and S. Van Buggenhout are researchers funded by the Research Foundation - Flanders (FWO). The authors would like to acknowledge ProMeta, the Interfacultary Centre for Proteomics and Metabolomics for the MALDI-TOF-MS analysis. Sincere thanks goes to Prof. J. Hofkens for giving us access to Olympus Fluoview 1000 confocal microscope.
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