Subcellular localization of the BtpA protein in the cyanobacterium Synechocystis sp. PCC 6803

Eur J Biochem. 1999 Apr;261(1):311-6. doi: 10.1046/j.1432-1327.1999.00281.x.

Abstract

Photosystem I is a large pigment-protein complex embedded in the thylakoid membranes of chloroplasts and cyanobacteria. In the cyanobacterium Synechocystis sp. PCC 6803, the btpA gene encodes a 30-kDa polypeptide. Mutations in this gene significantly affect accumulation of the reaction center proteins of photosystem I in Synechocystis 6803 [Bartsevich, V. V. & Pakrasi, H. B. (1997) J. Biol. Chem. 272, 6372-6378]. We describe here the intracellular localization of the BtpA protein. Immunolocalization in Synechocystis 6803 cells demonstrated that the BtpA protein is tightly associated with the thylakoid membranes. Phase fractionation in the detergent Triton X-114 indicated that BtpA is a peripheral membrane protein. To determine which surface of the thylakoid membrane BtpA is exposed to, we used a two-phase polymer partitioning technique to develop a novel method to isolate inside-out and right-side-out thylakoid vesicles from Synechocystis 6803. Treatments of such vesicles with different salts and protease showed that the BtpA protein is an extrinsic membrane protein which is exposed to the cytoplasmic face of the thylakoid membrane.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Chloroplasts / metabolism
  • Cyanobacteria / genetics
  • Cyanobacteria / metabolism*
  • Genes, Bacterial
  • Intracellular Membranes / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Photosynthetic Reaction Center Complex Proteins / metabolism
  • Subcellular Fractions / metabolism

Substances

  • Bacterial Proteins
  • BtpA protein, Synechocystis sp. PCC 6803
  • Membrane Proteins
  • Photosynthetic Reaction Center Complex Proteins