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Open Dataset XSBHEVPH

SEC-SAXS-MALS of Pseudomonas aeruginosa GluRS

Abstract

The aminoacyl-tRNA synthetases (AaRSs) are an ancient family of structurally diverse enzymes that deliver amino acids to the translation machinery. While GluRSs were previously classified as monomers, the form expressed in Pseudomonas aeruginosa appears to be a homodimer. This experiment was conducted to determine the oligomerization state in solution.

Experimental description

The SSGCID protein PsaeA.01348.a was treated with zinc to reconstitute the acceptor arm binding domain. The sample was diluted 7.5× with 20 mM HEPES, 150 mM NaCl, 5 mM MgCl2, pH 6.5, supplemented with 0.5 mM ZnSO4 and 10 mM TCEP, and then 2× with 20 mM HEPES, 150 mM NaCl, 5 mM MgCl2, pH 7.5, supplemented with 1 mM TCEP. The protein was flash frozen and stored at a final concentration of the approximately 3.5 mg/mL. At the beamline, the sample flowed through a Shodex 803 SEC column equilibrated with 20 mM HEPES, 150 mM NaCl, 5 mM MgCl2, pH 7.5, supplemented with 1 mM TCEP at 0.65 mL/min. The sample was exposed to X-rays of wavelength 1.127 Å continuously, with 2 sec exposure increments recorded with a Pilatus 3 2 M detector placed 2,100 mm away. The data set consisted of 660 intensity profiles, each spanning a q range of 0.0109 to 0.4729 Å−1.

File description

A_S_GluRS_1_5.dat

Created

2023-10-24

Published

2025-04-25

Data collection technique

SEC-SAXS

Journal DOI

Source

Advanced Light Source

Beamline

SIBYLS BL12.3.1

Wavelength

1.127 Å

Sample to Detector Distance

2.1 m

Submitting Author

Susan Tsutakawa

Lawrence Berkeley National Laboratory

United States of America

[email protected]

Collaborators

  • Isabelle Q Phan, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute
  • Michael K Fenwick, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute
  • Amy E Derocher, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute
  • Sandhya Subramanian, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute
  • Ariel B Abramov, [email protected], Seattle Structural Genomics Center for Infectious Disease, Seattle Children’s Research Institute
  • Ryan Choi, [email protected], Seattle Structural Genomics Center for Infectious Disease, Seattle Children’s Research Institute
  • Stephen N Hewitt, [email protected], Seattle Structural Genomics Center for Infectious Disease, Seattle Children’s Research Institute
  • Thomas E Edwards, [email protected], Seattle Structural Genomics Center for Infectious Disease, Seattle Children’s Research Institute, Union chimique belge (UCB)/Beryllium Discovery,
  • Iwona K Wower, [email protected], Department of Animal Sciences, Auburn University
  • Aimee C Soe, [email protected], Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory,
  • Jacek Wower, [email protected], Department of Animal Sciences, Auburn University
  • Bart L Staker, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute
  • Peter J Myler, [email protected], Seattle Structural Genomics Center for Infectious Disease and Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Department of Biomedical Information and Medical Education and Department of Global Health, University of Washington

Project Leader

Susan Tsutakawa

[email protected]


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Complete Set of SAS Data Files

The complete SAS dataset is downloadable as a zip file.

Individual SAS Data Files (Total: 1)

These are individual SAS data files that may be raw or processed (merged, etc.). These may or not be included in a zip file containing a larger dataset.

Supplemental Data and Supporting Materials (Total: 1)

These data and materials may include X-ray crystal structure coordinates, multi-angle light scattering data, .etc. It may also include additional details or methods pertaining to the SAXS experiments, or the researcher's interpretations of the results.

Open SAXS data analysis sites in a new tab using these links

SAXS Similarity SAXS FrameSlice

Sample:

  • Macromolecule 1: GluRS
    • Sample Full Name: Pseudomonas aeruginosa glutamyl tRNA synthetase
    • Sample Type: Protein
    • Source Organism: Pseudomonas aeruginosa
    • Source Organism NCBI Taxonomy ID:
    • Expression System: Escherichia coli BL21(DE3)-R3- pRARE2
    • Expression NCBI Taxonomy ID: 562
    • Uniprot ID: Q9XCL6
    • Sequence or Chemical Formula:
    • MAHHHHHHMTTVRTRIAPSPTGDPHVGTAYIALFNLCFARQHGGQFILRIEDTDQLRSTRESEQQIYDALRWLGIEWDEGPDVGGPHGPYRQSERGHIYKKYSDELVEKGHAFTCFCTPERLDAVRAEQMARKETPRYDGHCMHLPKDEVQRRLAAGESHVTRMKVPTEGVCVVPDMLRGDVEIPWDRMDMQVLMKADGLPTYFLANVVDDHLMGITHVLRGEEWLPSAPKLIKLYEYFGWEQPQLCYMPLLRNPDKSKLSKRKNPTSITFYERMGYLPQALLNYLGRMGWSMPDEREKFTLAEMIEHFDLSRVSLGGPIFDLEKLSWLNGQWIREQSVEEFAREVQKWALNPEYLMKIAPHVQGRVENFSQIAPLAGFFFSGGVPLDASLFEHKKLDPTQVRQVLQLVLWKLESLRQWEKERITGCIQAVAEHLQLKLRDVMPLMFPAITGHASSVSVLDAMEILGADLSRYRLRQALELLGGASKKETKEWEKIRDAIPG