Optimized native gel systems for separation of thylakoid protein complexes: novel super- and megacomplexes

Sari Sirpiö , Marjaana Suorsa , Virpi Paakkarinen , Eva-Mari Aro (1)

1 Department of Biochemistry and Food Chemistry

Gel-based analysis of thylakoid membrane protein complexes represents a valuable tool to monitor the dynamics of the photosynthetic machinery. The native polyacrylamide gel electrophoresis (native-PAGE) preserves the components and often also the conformation of the protein complexes, thus enabling the analysis of their subunit composition. Nevertheless, the literature and practical experimentation in the field sometimes raise confusion due to a great variety of native-PAGE and thylakoid solubilization systems. Here, we describe optimized methods for separation of higher plant thylakoid membrane protein complexes by native-PAGE addressing particularly (i) the use of detergent and (ii) solubilization buffer as well as (iii) the gel electrophoresis method. Special attention is paid on separation of high molecular mass thylakoid membrane super- and megacomplexes from Arabidopsis thaliana leaves. Several novel super- and megacomplexes including the photosystem (PS)I, PSII and light harvesting complexes (LHC) in various combinations are reported.

Mots clés: Life Sciences

Sari Sirpiö, Marjaana Suorsa, Virpi Paakkarinen, Eva-Mari Aro. Optimized native gel systems for separation of thylakoid protein complexes: novel super- and megacomplexes. Biochemical Journal, 2011, 439 (2), pp.207-214. ⟨10.1042/BJ20102155⟩. ⟨hal-00628664⟩