PDBsum entry 4dcs

Hydrolase

PDB id: 4dcs

Contents

Protein chain

406 a.a.

Ligands

GDP

SO4

Waters ×91

PDB id:

4dcs

Name:

Hydrolase

Title:

Crystal structure of b. Subtilis enga in complex with sulfate ion and gdp

Structure:

Gtp-binding protein enga. Chain: a. Synonym: gtp-binding protein yphc. Engineered: yes

Source:

Bacillus subtilis. Organism_taxid: 1423. Gene: der, enga, yphc, bsu22840. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.25Å R-factor: 0.219 R-free: 0.267

Authors:

J.-B.Reiser,D.Housset,A.-E.Foucher,J.-M.Jault

Key ref:

A.E.Foucher et al. (2012). Potassium acts as a GTPase-activating element on each nucleotide-binding domain of the essential Bacillus subtilis EngA. Plos One, 7, e46795. PubMed id: 23056455

Date:

18-Jan-12 Release date: 14-Nov-12

Protein chain

P50743  (DER_BACSU) -  GTPase Der from Bacillus subtilis (strain 168)

Seq: 436 a.a.

Struc: 406 a.a.

Enzyme reactions

• Enzyme class: E.C.?

Plos One 7:e46795 (2012)

PubMed id: 23056455

Potassium acts as a GTPase-activating element on each nucleotide-binding domain of the essential Bacillus subtilis EngA.

A.E.Foucher, J.B.Reiser, C.Ebel, D.Housset, J.M.Jault.

ABSTRACT

EngA proteins form a unique family of bacterial GTPases with two GTP-binding domains in tandem, namely GD1 and GD2, followed by a KH (K-homology) domain. They have been shown to interact with the bacterial ribosome and to be involved in its biogenesis. Most prokaryotic EngA possess a high GTPase activity in contrast to eukaryotic GTPases that act mainly as molecular switches. Here, we have purified and characterized the GTPase activity of the Bacillus subtilis EngA and two shortened EngA variants that only contain GD1 or GD2-KH. Interestingly, the GTPase activity of GD1 alone is similar to that of the whole EngA, whereas GD2-KH has a 150-fold lower GTPase activity. At physiological concentration, potassium strongly stimulates the GTPase activity of each protein construct. Interestingly, it affects neither the affinities for nucleotides nor the monomeric status of EngA or the GD1 domain. Thus, potassium likely acts as a chemical GTPase-activating element as proposed for another bacterial GTPase like MnmE. However, unlike MnmE, potassium does not promote dimerization of EngA. In addition, we solved two crystal structures of full-length EngA. One of them contained for the first time a GTP-like analogue bound to GD2 while GD1 was free. Surprisingly, its overall fold was similar to a previously solved structure with GDP bound to both sites. Our data indicate that a significant structural change must occur upon K(+) binding to GD2, and a comparison with T. maritima EngA and MnmE structures allowed us to propose a model explaining the chemical basis for the different GTPase activities of GD1 and GD2.