PDBsum entry 2gqc

Hydrolase

PDB id: 2gqc

Contents

Protein chain

70 a.a.

References listed in PDB file

Key reference

Title

Solution structure and dynamics of the n-Terminal cytosolic domain of rhomboid intramembrane protease from pseudomonas aeruginosa: insights into a functional role in intramembrane proteolysis.

Authors

A.Del rio, K.Dutta, J.Chavez, I.Ubarretxena-Belandia, R.Ghose.

Ref.

J Mol Biol, 2007, 365, 109-122. [DOI no: 10.1016/j.jmb.2006.09.047]

PubMed id

17059825

Abstract

Rhomboids are ubiquitous integral membrane proteases that release cellular signals from membrane-bound substrates through a general signal transduction mechanism known as regulated intramembrane proteolysis (RIP). We present the NMR structure of the cytosolic N-terminal domain (NRho) of P. aeruginosa Rhomboid. NRho consists of a novel alpha/beta fold and represents the first detailed structural insight into this class of intramembrane proteases. We find evidence that NRho is capable of strong and specific association with detergent micelles that mimic the membrane/water interface. Relaxation measurements on NRho reveal structural fluctuations on the microseconds-milliseconds timescale in regions including and contiguous to those implicated in membrane interaction. This structural plasticity may facilitate the ability of NRho to recognize and associate with membranes. We suggest that NRho plays a role in scissile peptide bond selectivity by optimally positioning the Rhomboid active site relative to the membrane plane.

Figure 1.
Figure 1. NRho forms a folded domain. (a) ^1H-^15N HSQC spectrum of NRho at 700 MHz displaying resonance assignments. The resonances corresponding to residues 82–87 were very weak and not assigned. (b) ^1H^N{^15N} NOE of NRho at 800 MHz. The broken red line indicates an NOE value of 0.25. Residues 1–70 were included in the structure calculation. Figure 1. NRho forms a folded domain. (a) ^1H-^15N HSQC spectrum of NRho at 700 MHz displaying resonance assignments. The resonances corresponding to residues 82–87 were very weak and not assigned. (b) ^1H^N{^15N} NOE of NRho at 800 MHz. The broken red line indicates an NOE value of 0.25. Residues 1–70 were included in the structure calculation.

Figure 6.
Figure 6. Membrane interaction site of NRho. (a) Key residues that interact with C[16]PN micelles lie in the β-sheet region of NRho and in the hairpin connecting β2 and β3. Residues that show attenuation in peak intensity in the presence of C[16]PN micelles for their backbone resonances are shaded red. Side-chains corresponding to Gln5 and Gln38 that also show peak attenuation in the presence of micelles are shown (green), as is the Trp41 side-chain (blue). (b) All residues that interact with C[16]PN micelles map onto a nearly continuous surface on one face of NRho. Key residues are labeled and the coloring scheme is the same as in (a). Figure 6. Membrane interaction site of NRho. (a) Key residues that interact with C[16]PN micelles lie in the β-sheet region of NRho and in the hairpin connecting β2 and β3. Residues that show attenuation in peak intensity in the presence of C[16]PN micelles for their backbone resonances are shaded red. Side-chains corresponding to Gln5 and Gln38 that also show peak attenuation in the presence of micelles are shown (green), as is the Trp41 side-chain (blue). (b) All residues that interact with C[16]PN micelles map onto a nearly continuous surface on one face of NRho. Key residues are labeled and the coloring scheme is the same as in (a).

The above figures are reprinted by permission from Elsevier: J Mol Biol (2007, 365, 109-122) copyright 2007.