Use of aminoglycoside adenyltransferase translational fusions to determine topology of thylakoid membrane proteins

John Lee Franklin, Jianying Zhang, Kevin Edward Redding

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have developed a system to examine the topology of thylakoid membrane proteins using the bacterial aadA gene as a reporter. Translational fusions that place the aminoglycoside adenyltransferase domain in the stroma should provide high antibiotic resistance, while those that place it in the thylakoid lumen should give rise to low resistance. Genes encoding chimeric polypeptides consisting of AadA fused to varying lengths of the PsaA polypeptide, whose topology is known, were introduced into the chloroplast genome of Chlamydomonas reinhardtii. As expected, chimeras with an even number of α-helices in general resulted in higher resistance. This effect was not due to differences in expression or in catalytic activity. This system should prove useful in analysis of novel proteins predicted to be localized to the thylakoid membrane.

Original languageEnglish
Pages (from-to)97-100
Number of pages4
JournalFEBS Letters
Volume536
Issue number1-3
DOIs
Publication statusPublished - Feb 11 2003

Fingerprint

Thylakoid Membrane Proteins
Thylakoids
Aminoglycosides
Fusion reactions
Genes
Topology
Chloroplast Genome
Chlamydomonas reinhardtii
Bacterial Genes
Peptides
Gene encoding
Microbial Drug Resistance
Catalyst activity
Anti-Bacterial Agents
Membranes
Proteins

Keywords

  • Aminoglycoside adenyltransferase
  • Chloroplast
  • Photosystem I
  • Thylakoid
  • Topology

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Use of aminoglycoside adenyltransferase translational fusions to determine topology of thylakoid membrane proteins. / Franklin, John Lee; Zhang, Jianying; Redding, Kevin Edward.

In: FEBS Letters, Vol. 536, No. 1-3, 11.02.2003, p. 97-100.

Research output: Contribution to journalArticle

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