The first α-helical domain of the vesicle-inducing protein in plastids 1 promotes oligomerization and lipid binding

Planta. 2013 Feb;237(2):529-40. doi: 10.1007/s00425-012-1772-1. Epub 2012 Oct 2.

Abstract

The vesicle-inducing protein in plastids 1 (Vipp1) is an essential component for thylakoid biogenesis in cyanobacteria and chloroplasts. Vipp1 proteins share significant structural similarity with their evolutionary ancestor PspA (bacterial phage shock protein A), namely a predominantly α-helical structure, the formation of oligomeric high molecular weight complexes (HMW-Cs) and a tight association with membranes. Here, we elucidated domains of Vipp1 from Arabidopsis thaliana involved in homo-oligomerization as well as association with chloroplast inner envelope membranes. We could show that the 21 N-terminal amino acids of Vipp1, which form the first α-helix of the protein, are essential for assembly of the 2 MDa HMW-C but are not needed for formation of smaller subcomplexes. Interestingly, removal of this domain also interferes with association of the Vipp1 protein to the inner envelope. Fourier transform infrared spectroscopy of recombinant Vipp1 further indicates that Escherichia coli lipids bind tightly enough that they can be co-purified with the protein. This feature also depends on the presence of the first helix, which strongly supports an interaction of lipids with the Vipp1 HMW-C but not with smaller subcomplexes. Therefore, Vipp1 oligomerization appears to be a prerequisite for its membrane association. Our results further highlight structural differences between Vipp1 and PspA, which might be important in regard to their different function in thylakoid biogenesis and bacterial stress response, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Bacterial Proteins / metabolism
  • Chloroplast Proteins / genetics
  • Chloroplast Proteins / metabolism
  • Chromatography, Gel
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Heat-Shock Proteins / metabolism
  • Intracellular Membranes / metabolism
  • Membrane Lipids / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Molecular Weight
  • Multiprotein Complexes / metabolism
  • Plastids / genetics
  • Plastids / metabolism*
  • Protein Binding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Spectroscopy, Fourier Transform Infrared

Substances

  • Arabidopsis Proteins
  • Bacterial Proteins
  • Chloroplast Proteins
  • Heat-Shock Proteins
  • Membrane Lipids
  • Membrane Proteins
  • Multiprotein Complexes
  • Recombinant Proteins
  • VIPP1 protein, Arabidopsis
  • phage shock protein, Bacteria