Qualitative und quantitative Untersuchung der Wechselwirkung des Immunodominanten Transmembran-Protein von ´Candidatus Phytoplasma mali´ mit Wirts-Aktin mittels Rasterkraftmikroskopie
Zusammenfassung der Projektergebnisse
`Ca. P.mali´ has only one major membrane protein, the immunodominant membrane protein (Imp), which is regarded as being close to the ancestor of all phytoplasma immunodominant membrane proteins. Imp binds to actin and possibly facilitates its movement in the plant or insect host cells. However, protein sequences of Imp are quite diverse among phytoplasma species, thus resulting in difficulties in identifying conserved domains across species. In this project, we combined physical and molecular biological methods to investigate an Imp-actin binding domain. We first compared the protein sequences of CaPmPM19 (Imp) with Imp of different strains of CaPm and identified 3 conserved domains (D1-3). Using different molecular biological methods we identified that D3 of Imp located at the carboxylic terminus of Imp is the actin binding domain. The D3 domain of Imp contains a highly positively charged amino acid cluster which is a common signature of actin binding proteins. Moreover we showed for the first time that Imp binds to actin of apple (Malus x domestica) which is the natural host plant of `Ca. P. mali`. The results of single molecule force spectroscopy analysis confirmed this finding. in the different adhesion force as well as the detachment work between Imp wild type, respectively D3 Imp mutant, and actin. The single molecule force spectroscopy could clearly show that domain D3 is involved in the specific actin-binding process of Imp. Thus our finding could lead to investigating a possible correlation between Imp variants and the infectivity of the corresponding `Ca. P. mali` isolates.
Projektbezogene Publikationen (Auswahl)
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Qualitative and quantitative investigation of the interaction of the immunodominant transmembrane protein from candidatus phytoplasma mali´ with host actin by Scanning Force Microscopy. ENFI 2018, Wittenberg
L. Kuaguim, C. Müller-Renno, M. Braun, K. Boonrod, G. Krczal, Ch. Ziegler
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Qualitative and quantitative investigation of the interaction of the immunodominant transmembrane protein from candidatus phytoplasma mali´with host actin by Scanning Force Microscopy. ENFI 2018, Wittenberg
L. Kuaguim, C. Müller-Renno, M. Braun, K. Boonrod, G. Krczal, Ch. Ziegler
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Rasterkraftmikroskopie zur qualitativen und quantitativen Untersuchung der Interaktion des immunodominanten Transmembranproteins von ´Candidatus Phytoplasma mali´ mit dem Wirtsaktin. AOFA 2018, Kaiserslautern
L. Kuaguim, C. Müller-Renno, M. Braun, K. Boonrod, G. Krczal, Ch. Ziegler
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Rasterkraftmikroskopie zur qualitativen und quantitativen Untersuchung der Interaktion des immunodominanten Transmembranproteins von ´Candidatus Phytoplasma mali´ mit dem Wirtsaktin. AOFA 2018, Kaiserslautern
L. Kuaguim, C. Müller-Renno, M. Braun, K. Boonrod, G. Krczal, Ch. Ziegler
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Single molecule force spectroscopy for qualitative and quantitative investigation of the interaction of the immunodominant transmembrane protein from ´Candidatus Phytoplasma mali´. Linz Winterworkshop 2019, Linz
L. Kuaguim, C. Müller-Renno, M. Braun, K. Boonrod, G. Krczal, Ch. Ziegler
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Identification of the Actin-Binding Region and Binding to Host Plant Apple Actin of Immunodominant Transmembrane Protein of ‘Candidatus Phytoplasma mali.’ International Journal of Molecular Sciences, 24(2), 968.
Boonrod, Kajohn; Kuaguim, Linda; Braun, Mario; Müller-Renno, Christine; Ziegler, Christiane & Krczal, Gabi
