Cellular localization of cytochrome bd in cyanobacteria using genetic code expansion

Mor Cohen, Eden Ozer, Ariel Kushmaro, Lital Alfonta

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Photosynthesis is one of the most fundamental and complex mechanisms in nature. It is a well-studied process, however, some photosynthetic mechanisms are yet to be deciphered. One of the many proteins that take part in photosynthesis, cytochrome bd, is a terminal oxidase protein that plays a role both in photosynthesis and in respiration in various organisms, specifically, in cyanobacteria. To clarify the role of cytochrome bd in cyanobacteria, a system for the incorporation of an unnatural amino acid into a genomic membrane protein cytochrome bd was constructed in Synechococcus sp. PCC7942. N-propargyl- l-lysine (PrK) was incorporated into mutants of cytochrome bd. Incorporation was verified and the functionality of the mutant cytochrome bd was tested, revealing that both electrochemical and biochemical activities were relatively similar to those of the wild-type protein. The incorporation of PrK was followed by a highly specific labeling and localization of the protein. PrK that was incorporated into the protein enabled a “click” reaction in a bio-orthogonal manner through its alkyne group in a highly specific manner. Cytochrome bd was found to be localized mostly in thylakoid membranes, as was confirmed by an enzyme-linked immunosorbent assay, indicating that our developed localization method is reliable and can be further used to label endogenous proteins in cyanobacteria.

Original languageEnglish
Pages (from-to)523-530
Number of pages8
JournalBiotechnology and Bioengineering
Issue number2
StatePublished - 1 Feb 2020


  • PCC 7942
  • cytochrome bd
  • electron transfer
  • genetic code expansion
  • site-specific labeling

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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