2L8Y

Solution structure of the E. coli outer membrane protein RcsF (periplasmatic domain)

Summary for 2L8Y

• Entry DOI: 10.2210/pdb2l8y/pdb
• NMR Information: BMRB: 17431
• Descriptor: Protein rcsF (1 entity in total)
• Functional Keywords: two component system, rcsf, rcs signalling system, disulphide bridges, exopolysaccharide biosynthesis, signaling protein
• Biological source: Escherichia coli
• Cellular location: Cell outer membrane: P69411
• Total number of polymer chains: 1
• Total formula weight: 11136.78

Authors

Rogov, V.V.,Rogova, N.Y.,Bernhard, F.,Lohr, F.,Doetsch, V. (deposition date: 2011-01-27, release date: 2011-04-06, Last modification date: 2024-10-30)

Primary citation

Rogov, V.V.,Rogova, N.Y.,Bernhard, F.,Lohr, F.,Dotsch, V.
A disulfide bridge network within the soluble periplasmic domain determines structure and function of the outer membrane protein RCSF.
J.Biol.Chem., 286:18775-18783, 2011

PubMed Abstract: RcsF, a proposed auxiliary regulator of the regulation of capsule synthesis (rcs) phosphorelay system, is a key element for understanding the RcsC-D-A/B signaling cascade, which is responsible for the regulation of more than 100 genes and is involved in cell division, motility, biofilm formation, and virulence. The RcsC-D-A/B system is one of the most complex bacterial signal transduction pathways, consisting of several membrane-bound and soluble proteins. RcsF is a lipoprotein attached to the outer membrane and plays an important role in activating the RcsC-d-A/B pathway. The exact mechanism of activation of the rcs phosphorelay by RcsF, however, remains unknown. We have analyzed the sequence of RcsF and identified three structural elements: 1) an N-terminal membrane-anchored helix (residues 3-13), 2) a loop (residues 14-48), and 3) a C-terminal folded domain (residues 49-134). We have determined the structure of this C-terminal domain and started to investigate its interaction with potential partners. Important features of its structure are two disulfide bridges between Cys-74 and Cys-118 and between Cys-109 and Cys-124. To evaluate the importance of this RcsF disulfide bridge network in vivo, we have examined the ability of the full-length protein and of specific Cys mutants to initiate the rcs signaling cascade. The results indicate that the Cys-74/Cys-118 and the Cys-109/Cys-124 residues correlate pairwise with the activity of RcsF. Interaction studies showed a weak interaction with an RNA hairpin. However, no interaction could be detected with reagents that are believed to activate the rcs phosphorelay, such as lysozyme, glucose, or Zn(2+) ions.

PubMed: 21471196
DOI: 10.1074/jbc.M111.230185

Experimental method

SOLUTION NMR