Application of split-green fluorescent protein for topology mapping membrane proteins in Escherichia coli
2012 (English)In: Protein Science, ISSN 0961-8368, E-ISSN 1469-896X, Vol. 21, no 10, 1571-1576 p.Article in journal (Refereed) Published
A topology map of a membrane protein defines the location of transmembrane helices and the orientation of soluble domains relative to the membrane. In the absence of a high-resolution structure, a topology map is an essential guide for studying structurefunction relationships. Although these maps can be predicted directly from amino acid sequence, the predictions are more accurate if combined with experimental data, which are usually obtained by fusing a reporter protein to the C-terminus of the protein. However, as reporter proteins are large, they cannot be used to report on the cytoplasmic/periplasmic location of the N-terminus of a protein. Here, we show that the bimolecular split-green fluorescent protein complementation system can overcome this limitation and can be used to determine the location of both the N- and C-termini of inner membrane proteins in Escherichia coli.
Place, publisher, year, edition, pages
2012. Vol. 21, no 10, 1571-1576 p.
topology, membrane protein, split-GFP, inner membrane
Biochemistry and Molecular Biology
Research subject Biochemistry
IdentifiersURN: urn:nbn:se:su:diva-82120DOI: 10.1002/pro.2131ISI: 000308994300015OAI: oai:DiVA.org:su-82120DiVA: diva2:566757