Class III peroxidases: Functional analysis of membrane-bound peroxidases
Final Report Abstract
At least three peroxidases have been identified in highly enriched plasma membrane fractions of maize (Zea mays L.) roots by our team. Membrane localization of these proteins was verified by different methods (GFP fusion proteins, Immuno-gold labelling). Using real time qPCR distribution of transcripts was analyzed in different root zones. A differential regulation of the genes has been shown by treatment with several stressors. Expression of ZmPrx01 was mostly affected by Chitosan. ZmPrx66 was mostly affected by H2O2 and ZmPrx70 was mostly affected by salicylic acid. Compared to our former data on protein abundance salicylic acid showed a well correlation. Abundance of Prx increased by chitosan and wounding. In contrast to this result ZmPrx66 decreased by H2O2 and wounding. Other peroxidases increased by treatment with these effectors. Cadmium stress was investigated in more detail (short term and long term). ZmPrx01 was not affected by cadmium over short term, whereas a decrease of the transcript was observed for long term experiments. ZmPrx66 and ZmPrx70 were up-regulated within 15 to 30 minutes after Cadmium exposure. This result suggests a co-regulation with RbohD. Long term experiments showed a down-regulation of ZmPrx66 and ZmPrx70. Several proteomic approaches have been established for analysis of soluble and membranebound peroxidases. Evidence for a location of these proteins in detergent insoluble membranes and high molecular mass protein complexes has been published. Functional analysis of the peroxidases has been done by heterologous expression of the peroxidases in Pichia pastoris. Biochemical characterization has been performed for the recombinant Prx70. In general results of the recombinant Protein were comparable with the native protein. The recombinant protein, however, showed a higher termostability, which was possibly due to the higher glycosylation in Pichia. Ferulic acid, coumaric acid and coniferyl alcohol were used as substrates by Prx70rec. Besides biochemical characterization RNAi mutants have been produced for all three peroxidases. Until now there are more than 130 new developed maize strains not suitable analysed, yet.
Publications
- (2011) Phylogeny, topology, structure and functions of membrane-bound class III peroxidases in vascular plants. Phytochemistry 72:1124-1135
S. Lüthje, C.N. Meisrimler, D. Hopff, B. Möller
- (2012) Cell wall-bound cationic and anionic class III isoperoxidases of pea root: biochemical characterization and function in root growth. Journal of Experimental Botany 63:4631-45.
B.M. Kukavica , S.D. Veljovic-Jovanovic, Lj. Menckhoff, S. Lüthje
(See online at https://doi.org/10.1093/jxb/ers139) - (2013) Redox pathways in plant plasma membranes and oxidative stress. Antioxidants and Redox Signaling 18:2163-83
S. Lüthje, B. Möller, F.C. Perrineau, K. Wöltje
- (2014) Alterations in Soluble Class III Peroxidases of Maize Shoots by Flooding Stress. Proteomes 2 (3), 303-322
C.N. Meisrimler, F. Buck, S. Lüthje
(See online at https://dx.doi.org/10.3390/proteomes2030303) - (2014) Class III peroxidases. In: Methods in Molecular Plant Biology, Plant Proteomics: Methods and Protocols, Second Edition, eds. J.V. Jorrín Novo, S. Komatsu, W. Weckwerth, S. Wienkoop; pp. 687-706, New York, NY: Humana Press Inc.
S. Lüthje, C.N. Meisrimler, D. Hopff, T. Schütze, Jenny Köppe, K. Heino
- (2015) Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining. Front. Plant Sci.
C.N. Meisrimler, A. Schwendke, S. Lüthje
(See online at https://doi.org/10.3389/fpls.2015.00230)
