2L5H

Solution Structure of the H189Q mutant of the Enzyme I dimer Using Residual Dipolar Couplings and Small Angle X-Ray Scattering

Summary for 2L5H

• Entry DOI: 10.2210/pdb2l5h/pdb
• Related: 2HRO 2HWG 2KX9 2WQD 2XDF 3EZA
• Descriptor: Phosphoenolpyruvate-protein phosphotransferase (1 entity in total)
• Functional Keywords: protein, dimer, transferase
• Biological source: Escherichia coli
• Cellular location: Cytoplasm: P08839
• Total number of polymer chains: 2
• Total formula weight: 126818.66

Authors

Takayama, Y.D.,Schwieters, C.D.,Grishaev, A.,Guirlando, R.,Clore, G. (deposition date: 2010-11-01, release date: 2011-01-12, Last modification date: 2024-05-01)

Primary citation

Takayama, Y.,Schwieters, C.D.,Grishaev, A.,Ghirlando, R.,Clore, G.M.
Combined Use of Residual Dipolar Couplings and Solution X-ray Scattering To Rapidly Probe Rigid-Body Conformational Transitions in a Non-phosphorylatable Active-Site Mutant of the 128 kDa Enzyme I Dimer.
J.Am.Chem.Soc., 133:424-427, 2011

PubMed Abstract: The first component of the bacterial phosphotransferase system, enzyme I (EI), is a multidomain 128 kDa dimer that undergoes large rigid-body conformational transitions during the course of its catalytic cycle. Here we investigate the solution structure of a non-phosphorylatable active-site mutant in which the active-site histidine is substituted by glutamine. We show that perturbations in the relative orientations and positions of the domains and subdomains can be rapidly and reliably determined by conjoined rigid-body/torsion angle/Cartesian simulated annealing calculations driven by orientational restraints from residual dipolar couplings and shape and translation information afforded by small- and wide-angle X-ray scattering. Although histidine and glutamine are isosteric, the conformational space available to a Gln side chain is larger than that for the imidazole ring of His. An additional hydrogen bond between the side chain of Gln189 located on the EIN(α/β) subdomain and an aspartate (Asp129) on the EIN(α) subdomain results in a small (∼9°) reorientation of the EIN(α) and EIN(α/β) subdomains that is in turn propagated to a larger reorientation (∼26°) of the EIN domain relative to the EIC dimerization domain, illustrating the positional sensitivity of the EIN domain and its constituent subdomains to small structural perturbations.

PubMed: 21162528
DOI: 10.1021/ja109866w

Experimental method

SOLUTION NMR
SOLUTION SCATTERING