Multiprotein Complexes Governing Chlorophyll Biosynthesis

• Applicant: Dr. Dmitry Shvarev
• Subject Area: Biochemistry
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 537976353

The process of chlorophyll biogenesis is complex and involves intricate multiprotein complexes. However, the mechanistic understanding of the underlying enzymatic reactions is poor due to a lack of structural information. The aim of this project is to study the multiprotein complexes involved in chlorophyll biosynthesis and to gain mechanistic insights into this process. Structural biology techniques such as cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) will be used together with biochemical characterization and fluorescence microscopy localization methods. The study will focus on three objectives. First, the magnesium chelatase enzyme complex will be studied by reconstitution and determination of its cryo-EM structures, its conformational dynamics will be analyzed, and its function will be elucidated by enzymatic assays and cellular localization. In addition, the role of proteins interacting with magnesium chelatase and the regulation of the complex will be investigated by determining the structures of the complex with binding partners by cryo-EM, accompanied by biochemical and localization analyses. Finally, the last step of chlorophyll biosynthesis, the insertion of chlorophyll into chlorophyll-containing proteins, will be investigated by cryo-EM analysis, biochemical and localization studies, and cryo-ET to understand the structural organization of chlorophyll incorporation. The results of this project will advance the understanding of chlorophyll biosynthesis and photosynthesis. High-resolution structural information on chlorophyll-synthesizing complexes could support future research and applications in areas such as agriculture and biotechnology, among others.

• DFG Programme: Independent Junior Research Groups
• Major Instrumentation: Liquid chromatography system
• Instrumentation Group: 1350 Flüssigkeits-Chromatographen (außer Aminosäureanalysatoren 317), Ionenaustauscher