Characterization of lacZ complementation deletions using membrane receptor dimerization

D. Thormeyer, O. Ammerpohl, O. Larsson, Y. Xu, A. Asinger, Claes R Wahlestedt, Zicai Liang

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We describe the screening of lacZ deletions in mammalian cells and the discovery of a novel pair of lacZ deletions that can undergo α-complementation only when they are fused to peptides that interact with each other. The two lacZ deletions, ΔN 11-75 and ΔC 82-1023, were first characterized by fusing to two small interacting peptides and were then further analyzed by fusing to three membrane receptors (G protein-coupled receptors α2cAR and D2DRL and receptor tyrosine kinase insulin receptor) that were known to form homodimers in the membrane. Histochemical and quantitative FACS® assays demonstrated that the novel deletions have much lower level of association with each other thus offering a much lower background in monitoring membrane protein interactions compared to previously published lacZ deletions. Furthermore, our method has the exciting potential to monitor simultaneously membrane receptor dimerization and localization to the cell surface of living cells.

Original languageEnglish
Pages (from-to)346-355
Number of pages10
JournalBioTechniques
Volume34
Issue number2
StatePublished - Feb 1 2003
Externally publishedYes

Fingerprint

Dimerization
Membranes
Cells
Peptides
Insulin Receptor
Receptor Protein-Tyrosine Kinases
G-Protein-Coupled Receptors
Assays
Screening
Membrane Proteins
Association reactions
Monitoring

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Thormeyer, D., Ammerpohl, O., Larsson, O., Xu, Y., Asinger, A., Wahlestedt, C. R., & Liang, Z. (2003). Characterization of lacZ complementation deletions using membrane receptor dimerization. BioTechniques, 34(2), 346-355.

Characterization of lacZ complementation deletions using membrane receptor dimerization. / Thormeyer, D.; Ammerpohl, O.; Larsson, O.; Xu, Y.; Asinger, A.; Wahlestedt, Claes R; Liang, Zicai.

In: BioTechniques, Vol. 34, No. 2, 01.02.2003, p. 346-355.

Research output: Contribution to journalArticle

Thormeyer, D, Ammerpohl, O, Larsson, O, Xu, Y, Asinger, A, Wahlestedt, CR & Liang, Z 2003, 'Characterization of lacZ complementation deletions using membrane receptor dimerization', BioTechniques, vol. 34, no. 2, pp. 346-355.
Thormeyer D, Ammerpohl O, Larsson O, Xu Y, Asinger A, Wahlestedt CR et al. Characterization of lacZ complementation deletions using membrane receptor dimerization. BioTechniques. 2003 Feb 1;34(2):346-355.
Thormeyer, D. ; Ammerpohl, O. ; Larsson, O. ; Xu, Y. ; Asinger, A. ; Wahlestedt, Claes R ; Liang, Zicai. / Characterization of lacZ complementation deletions using membrane receptor dimerization. In: BioTechniques. 2003 ; Vol. 34, No. 2. pp. 346-355.
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