NDLI: Topological analysis of the Escherichia coli WcaJ protein reveals a new conserved configuration for the polyisoprenyl-phosphate hexose-1-phosphate transferase family

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Topological analysis of the Escherichia coli WcaJ protein reveals a new conserved configuration for the polyisoprenyl-phosphate hexose-1-phosphate transferase family

Content Provider	Semantic Scholar
Author	Furlong, Sarah E. Ford, Amy M. Albarnez-Rodriguez, Lorena Valvano, Miguel A.
Copyright Year	2015
Abstract	WcaJ is an Escherichia coli membrane enzyme catalysing the biosynthesis of undecaprenyl-diphosphate-glucose, the first step in the assembly of colanic acid exopolysaccharide. WcaJ belongs to a large family of polyisoprenyl-phosphate hexose-1-phosphate transferases (PHPTs) sharing a similar predicted topology consisting of an N-terminal domain containing four transmembrane helices (TMHs), a large central periplasmic loop, and a C-terminal domain containing the fifth TMH (TMH-V) and a cytosolic tail. However, the topology of PHPTs has not been experimentally validated. Here, we investigated the topology of WcaJ using a combination of LacZ/PhoA reporter fusions and sulfhydryl labelling by PEGylation of novel cysteine residues introduced into a cysteine-less WcaJ. The results showed that the large central loop and the C-terminal tail both reside in the cytoplasm and are separated by TMH-V, which does not fully span the membrane, likely forming a "hairpin" structure. Modelling of TMH-V revealed that a highly conserved proline might contribute to a helix-break-helix structure in all PHPT members. Bioinformatic analyses show that all of these features are conserved in PHPT homologues from Gram-negative and Gram-positive bacteria. Our data demonstrate a novel topological configuration for PHPTs, which is proposed as a signature for all members of this enzyme family.
File Format	PDF HTM / HTML
DOI	10.1038/srep09178
PubMed reference number	25776537
Journal	Medline
Volume Number	5
Alternate Webpage(s)	https://pure.qub.ac.uk/portal/files/14413861/Furlong_15_SREP.pdf
Alternate Webpage(s)	http://pure.qub.ac.uk/portal/files/14413861/Furlong_15_SREP.pdf
Alternate Webpage(s)	https://pureadmin.qub.ac.uk/ws/portalfiles/portal/14413861/Furlong_15_SREP.pdf
Journal	Scientific reports
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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