2022 Membrane destabilization and pore formation induced by the Synechocystis IM30 protein

Benedikt Junglas1, Amelie Axt23, Carmen Siebenaller1, Hilal Sonel1, Nadja Hellmann1, Stefan A.L. Weber23, Dirk Schneider14

Biophysical Journals, Volume 121, Issue 18, 20 September 2022, Pages 3411-3421

1 Department of Chemistry, Biochemistry, Johannes Gutenberg University Mainz, Mainz, Germany
2 Max Planck-Institute for Polymer Research, Mainz, Germany
3 Institute of Physics, Johannes Gutenberg University Mainz, Mainz, Germany
4 Institute of Molecular Physiology, Johannes Gutenberg University Mainz, Mainz, Germany

The inner membrane-associated protein of 30 kDa (IM30) is essential in chloroplasts and cyanobacteria. The spatio-temporal cellular localization of the protein appears to be highly dynamic and triggered by internal as well as external stimuli, mainly light intensity. The soluble fraction of the protein is localized in the cyanobacterial cytoplasm or the chloroplast stroma, respectively. Additionally, the protein attaches to the thylakoid membrane as well as to the chloroplast inner envelope or the cyanobacterial cytoplasmic membrane, respectively, especially under conditions of membrane stress. IM30 is involved in thylakoid membrane biogenesis and/or maintenance, where it either stabilizes membranes and/or triggers membrane-fusion processes. These apparently contradicting functions have to be tightly controlled and separated spatiotemporally in chloroplasts and cyanobacteria. IM30’s fusogenic activity depends on Mg2+ binding to IM30; yet, it still is unclear how Mg2+-loaded IM30 interacts with membranes and promotes membrane fusion. Here, we show that the interaction of Mg2+ with IM30 results in increased binding of IM30 to native, as well as model, membranes. Via atomic force microscopy in liquid, IM30-induced bilayer defects were observed in solid-supported bilayers in the presence of Mg2+. These structures differ dramatically from the membrane-stabilizing carpet structures that were previously observed in the absence of Mg2+. Thus, Mg2+-induced alterations of the IM30 structure switch the IM30 activity from a membrane-stabilizing to a membrane-destabilizing function, a crucial step in membrane fusion.