Oct 29, 2018

Cell-free soluble expression of the membrane protein PsbS

BioRxiv : the Preprint Server for Biology
M KrishnanA Pandit

Abstract

Photosystem II subunit S (PsbS) is a membrane protein that plays an exclusive role in non-photochemical quenching for photoprotection of plants under high-light conditions. The activation mechanism of PsbS and its pH-induced conformational changes are currently unknown. For structural investigation of PsbS, effective synthesis of PsbS with selective isotope or electron-spin labels or non-natural amino acids incorporated would be a great asset. This communication presents cell-free expression as a successful method for in vitro production of PsbS that would allow such incorporation. We have optimized the cell-free method to yield soluble PsbS of ~500 ng/microl using a continuous-exchange method at 30 0C, along with a successful purification and refolding of PsbS in n-Dodecyl b-D-maltoside (b-DM) detergent. We expect that the presented protocols are transferrable for in vitro expression of other membrane proteins of the Light-Harvesting Complex family.

  • References
  • Citations

References

  • We're still populating references for this paper, please check back later.
  • References
  • Citations

Citations

  • This paper may not have been cited yet.

Mentioned in this Paper

Solocyte
Photosystem I, PSI-G subunit
Amino Acids, I.V. solution additive
Molecular Probe Techniques
Plasma Exchange
Evaluation
Soluble
Electron Microscopy
Purification Aspects
Membrane Proteins

About this Paper

Related Feeds

BioRxiv & MedRxiv Preprints

BioRxiv and MedRxiv are the preprint servers for biology and health sciences respectively, operated by Cold Spring Harbor Laboratory. Here are the latest preprint articles (which are not peer-reviewed) from BioRxiv and MedRxiv.