the hetero-hexameric nature of a chloroplast aaa+ ftsh protease contributes to its thermodynamic stability叶绿体的hetero-hexameric自然aaa + ftsh蛋白酶有助于其热力学稳定.pdfVIP

The Hetero-Hexameric Nature of a Chloroplast AAA+ FtsH Protease Contributes to Its Thermodynamic Stability 1. 2. 3 1 2 Ofer Moldavski , Olga Levin-Kravets , Tamar Ziv , Zach Adam , Gali Prag * 1The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Hebrew University of Jerusalem, Rehovot, Israel, 2 Department of Biochemistry and Molecular Biology and the Institute for Structural Biology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel, 3 Department of Biology, Smoler Proteomics Center, Technion, Haifa, Israel Abstract FtsH is an evolutionary conserved membrane-bound metalloprotease complex. While in most prokaryotes FtsH is encoded by a single gene, multiple FtsH genes are found in eukaryotes. Genetic and biochemical data suggest that the Arabidopsis chloroplast FtsH is a hetero-hexamer. This raises the question why photosynthetic organisms require a heteromeric complex, whereas in most bacteria a homomeric one is sufficient. To gain structural information of the possible complexes, the Arabidopsis FtsH2 (type B) and FtsH5 (type A) were modeled. An in silico study with mixed models of FtsH2/5 suggests that heteromeric hexamer structure with ratio of 4:2 is more likely to exists. Specifically, calculation of the buried surface area at the interfaces between neighboring subunits revealed that a hetero-complex should be thermodynamically more stable than a homo-hexamer, due to the presence of additional hydrophobic and hydrophilic interactions. To biochemically assess this model, we generated Arabidopsis transgenic plants, expressing epitope-tagged FtsH2 and immuno-purified the protein. Mass-spectrometry analysis showed that FtsH2 is associated