Application of split-green fluorescent protein for topology mapping membrane proteins in Escherichia coli |
| |
Authors: | Stephen Toddo Bill Söderström Isolde Palombo Gunnar von Heijne Morten H H Nørholm Daniel O Daley |
| |
Affiliation: | Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, SE-106 91 Sweden. |
| |
Abstract: | 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 structure-function 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. |
| |
Keywords: | topology membrane protein split‐GFP inner membrane |
本文献已被 PubMed 等数据库收录! |
|