Use of phoA fusions to study the topology of the Escherichia coli inner membrane protein leader peptidase.
AUTOR(ES)
San Millan, J L
RESUMO
A topology of the Escherichia coli leader peptidase has been previously proposed on the basis of proteolytic studies. Here, a collection of alkaline phosphatase fusions to leader peptidase is described. Fusions to the periplasmic domain of this protein exhibit high alkaline phosphatase activity, while fusions to the cytoplasmic domain exhibit low activity. Elements within the cytoplasmic domain are necessary to stably anchor alkaline phosphatase in the cytoplasm. The amino-terminal hydrophobic segment of leader peptidase acts as a weak export signal for alkaline phosphatase. However, when this segment is preceded by four lysines, it acts as a highly efficient export signal. The coherence of in vitro studies with alkaline phosphatase fusion analysis of the topology of leader peptidase further indicates the utility of this genetic approach to membrane protein structure and insertion.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=210394Documentos Relacionados
- Analysis of Escherichia coli TonB membrane topology by use of PhoA fusions.
- Use of phoA and lacZ fusions to study the membrane topology of ProW, a component of the osmoregulated ProU transport system of Escherichia coli.
- Use of gene fusions to determine the orientation of gene phoA on the Escherichia coli chromosome.
- Membrane topology model of Escherichia coli alpha-ketoglutarate permease by phoA fusion analysis.
- A plasmid facilitating in vitro construction of phoA gene fusions in Escherichia coli.