SASDAX6

Protein CyaY monomer

Data type: SASBDB experimental data Status: Published Curve type: Single concentration Last updated: 2023-05-25T11:21:52.654103+02:00

Reduced levels of frataxin, an essential protein of as yet unknown function, are responsible for causing the neurodegenerative pathology Friedreich’s ataxia. Independent reports have linked frataxin to iron–sulphur cluster assembly through interactions with the two central components of this machinery: desulphurase Nfs1/IscS and the scaffold protein Isu/IscU. In this study, we use a combination of biophysical methods to define the structural bases of the interaction of CyaY (the bacterial orthologue of frataxin) with the IscS/IscU complex. We show that CyaY binds IscS as a monomer in a pocket between the active site and the IscS dimer interface. Recognition does not require iron and occurs through electrostatic interactions of complementary charged residues. Mutations at the complex interface affect the rates of enzymatic cluster formation. CyaY binding strengthens the affinity of the IscS/IscU complex. Our data suggest a new paradigm for understanding the role of frataxin as a regulator of IscS functions.

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1. Samples, Components and Experimental Conditions Sample & Experiment

Sample 1 · Protein CyaY monomer

Concentration— – 5.0 mg/mlBuffer / pH20 mM Tris-HCl 150 mM NaCl 10 mM β-mercaptoethanol / 8.0
Experimental temperature15.0 Facility / beamlineDORIS III, DESY / EMBL X33
Wavelength0.15 nmExposure30.0 s × 4

Molecular Components

ComponentType / organismUniProt and ConstructOligomeric StateMolecular weight
Protein CyaY
View sequence
MNDSEFHRLADQLWLTIEERLDDWDGDSDIDCEINGGVLTITFENGSKIIINRQEPLHQVWLATKQGGYHFDLKGDEWICDRSGETFWDLLEQAATQQAGETVSFR
proteinEscherichia coliP27838—–—monomerMolecule count 112.231 kDa

Experimental Curves

CurvePoints / columnsq rangeUncertainty qualityNegative-intensity pointsSource file
12310[3]0.0774543–6.23701 1/nmHas uncertainty columnMissing 027sasbdb/entries/x6/sasdax6/source/SASDAX6.dat
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2. Measurements Reported by SASBDB Reported Results

MetricMethodValueUncertaintyUnit
dmaxP(r)5.0—nm
i0Guinier14.8——
i0P(r)14.8——
mwExperimental11.0—kDa
mwGuinier I(0)11.0—kDa
rgGuinier1.530.004nm
rgP(r)1.53—nm

These values are reported by SASBDB; SAXSdb did not recalculate them from the experimental curve.

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3. Source Fits and Models Source Fits & Models

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4. Source File Index Source Files

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5. Experiment Description and Publication Experiment & Publication

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6. Complete Source Record Complete Source Record

The following content comes directly from the SASBDB entry. Missing values are shown as “—”; SAXSdb does not infer undeclared units.

Open the original SASBDB entry

Buffer and Sample Properties

Buffer name20 mM Tris-HCl 150 mM NaCl 10 mM β-mercaptoethanolBuffer concentration20.0 mM
pH8.0Additives150 mM NaCl and 10 mM β-mercaptoethanol
Buffer description—
Purity measurement method—Extinction coefficient—
Absorbance—Scattering contrast—
Specific volume / dry volume— / —Mixture / deuteration— / —

Acquisition Conditions and Instrument

Measurement date2009-11-05Storage / measurement temperature15.0 / 15.0
Exposure time30.0Frames4
Wavelength0.15Sample-to-detector distance2.7
SourceX-ray synchrotronDetectorPilatus 1M-W
Facility / beamlineDORIS III, DESY / EMBL X33 · Hamburg, Germany
q range0.077 – 6.237Sample volume / flow rate— / —

Original SASBDB Plots

Experimental I(q)
Experimental I(q)
Experimental I(q), log-log
Experimental I(q), log-log
Guinier plot
Guinier plot
Kratky plot
Kratky plot
P(r) plot
P(r) plot

Downloadable Files

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full_entry_zip:source record and ZIP contents (6 items)
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pddf:source record
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All source fields (unfiltered)

Every source field is expanded here, including empty values, list members and fields already shown above.

summary.json:182 field values
Field pathRaw value
codeSASDAX6
statusPublished
type_of_curveSingle concentration
angular_unit1/nm
project.titleStructural bases for the interaction of frataxin with the central components of iron-sulphur cluster assembly.
project.publication.titleStructural bases for the interaction of frataxin with the central components of iron-sulphur cluster assembly.
project.publication.author_listPrischi F, Konarev PV, Iannuzzi C, Pastore C, Adinolfi S, Martin SR, Svergun DI, Pastore A
project.publication.journalNat Commun
project.publication.doi10.1038/ncomms1097
project.publication.pmid20981023
project.publication.published_date2010 Oct 19
project.statusreleased
project.submitted_datenull
project.released_date2015-07-07
pddf_datahttps://www.sasbdb.org/media/p_of_R_files/SASDAX6.out
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intensities_guinier_plothttps://www.sasbdb.org/media/intensities_files/scattering_plots/SASDAX6_guinier_img.png
sascif_datahttps://www.sasbdb.org/media/sascif/sascif_files/SASDAX6.sascif
experiment.instrument.detector.typeCustom-built
experiment.instrument.detector.namePilatus 1M-W
experiment.instrument.detector.resolution0.172
experiment.instrument.nameDORIS III, DESY
experiment.instrument.cityHamburg
experiment.instrument.countryGermany
experiment.instrument.beamline_nameEMBL X33
experiment.instrument.beam_geometry0.60
experiment.instrument.type_of_sourceX-ray synchrotron
experiment.instrument.point_sourcenull
experiment.instrument.line_collimationnull
experiment.instrument.sample_path_lengthnull
experiment.instrument.line_collimation_slitlengthnull
experiment.instrument.line_collimation_integrationwidthnull
experiment.instrument.xray_energynull
experiment.instrument.beam_profile_ahnull
experiment.instrument.beam_profile_alnull
experiment.sample.molecule[0].long_nameProtein CyaY
experiment.sample.molecule[0].short_namenull
experiment.sample.molecule[0].sequenceMNDSEFHRLADQLWLTIEERLDDWDGDSDIDCEINGGVLTITFENGSKIIINRQEPLHQVWLATKQGGYHFDLKGDEWIC DRSGETFWDLLEQAATQQAGETVSFR
experiment.sample.molecule[0].organismEscherichia coli
experiment.sample.molecule[0].uniprot_codeP27838
experiment.sample.molecule[0].uniprot_range_firstnull
experiment.sample.molecule[0].uniprot_range_lastnull
experiment.sample.molecule[0].oligomerizationmonomer
experiment.sample.molecule[0].molecular_typeprotein
experiment.sample.molecule[0].uniprot_sequenceMNDSEFHRLADQLWLTIEERLDDWDGDSDIDCEINGGVLTITFENGSKIIINRQEPLHQV WLATKQGGYHFDLKGDEWICDRSGETFWDLLEQAATQQAGETVSFR
experiment.sample.molecule[0].mw12.231
experiment.sample.molecule[0].total_mw12.231
experiment.sample.molecule[0].number_molecules1
experiment.sample.molecule[0].complex_statenull
experiment.sample.molecule[0].deuterationnull
experiment.sample.molecule[0].molecule_sourcebiological
experiment.sample.molecule[0].molecule_descriptionnull
experiment.sample.buffer.name20 mM Tris-HCl 150 mM NaCl 10 mM β-mercaptoethanol
experiment.sample.buffer.concentration_unitmM
experiment.sample.buffer.commentnull
experiment.sample.buffer.additive150 mM NaCl and 10 mM β-mercaptoethanol
experiment.sample.buffer.concentration20.0
experiment.sample.buffer.pkanull
experiment.sample.buffer.ph8.0
experiment.sample.buffer.deuterationnull
experiment.sample.purity_methodnull
experiment.sample.nameProtein CyaY monomer
experiment.sample.ext_coefficientnull
experiment.sample.contrastnull
experiment.sample.specific_volnull
experiment.sample.dry_volnull
experiment.sample.absorbptionnull
experiment.sample.deuterationnull
experiment.sample.mixturenull
experiment.contributor[0].affiliation[0].short_nameEMBL-Hamburg
experiment.contributor[0].affiliation[0].addressNotkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany
experiment.contributor[0].affiliation[0].full_nameEuropean Molecular Biology Laboratory (EMBL) - Hamburg outstation
experiment.contributor[0].affiliation[0].webpagehttp://www.embl-hamburg.de/index.php
experiment.contributor[0].contributor_namePetr
experiment.contributor[0].contributor_surnameKonarev
experiment.contributor[0].orcidnull
experiment.concentration_methodnull
experiment.concentration_unitmg/ml
experiment.date2009-11-05
experiment.storage_temperature15.0
experiment.cell_temperature15.0
experiment.exposure_time30.0
experiment.number_of_frames4
experiment.wavelength0.15
experiment.sample_detector_distance2.7
experiment.concentration_minnull
experiment.concentration_max5.0
experiment.sample_volumenull
experiment.flow_ratenull
experiment.s_min0.077
experiment.s_max6.237
experiment.total_exposure_timenull
experiment.seccolumnnull
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fits[0].models[0].softwarePDB (Protein Data Bank)
fits[0].models[0].pdb_link[0].pdb_code1soy
fits[0].models[0].pdb_link[0].ext_code1SOY
fits[0].models[0].pdb_link[0].ext_typepdb
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fits[0].models[0].model_mw12.22
fits[0].models[0].bead_radius1.9
fits[0].models[0].lognull
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fits[0].models[0].comment
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fits[0].softwarenull
fits[0].chi_square_value1.447209
fits[0].p_value0.0
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fits[1].p_value0.351246
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fits[1].descriptionnull
estimated_volume_methodnull
pddf_softwareATSAS GNOM
pddf_software_versionGNOM
i0_calibration_standardbovin serum albumin
descriptionReduced levels of frataxin, an essential protein of as yet unknown function, are responsible for causing the neurodegenerative pathology Friedreich’s ataxia. Independent reports have linked frataxin to iron–sulphur cluster assembly through interactions with the two central components of this machinery: desulphurase Nfs1/IscS and the scaffold protein Isu/IscU. In this study, we use a combination of biophysical methods to define the structural bases of the interaction of CyaY (the bacterial orthologue of frataxin) with the IscS/IscU complex. We show that CyaY binds IscS as a monomer in a pocket between the active site and the IscS dimer interface. Recognition does not require iron and occurs through electrostatic interactions of complementary charged residues. Mutations at the complex interface affect the rates of enzymatic cluster formation. CyaY binding strengthens the affinity of the IscS/IscU complex. Our data suggest a new paradigm for understanding the role of frataxin as a regulator of IscS functions.
experiment_descriptionnull
tags[0]X33
intensity_unitnull
experimental_mw11.0
experimental_mw_errornull
guinier_i0_mw11.0
guinier_i0_mw_errornull
porod_mwnull
porod_mw_errornull
pddf_i014.8
pddf_i0_errornull
guinier_i014.8
guinier_i0_errornull
pddf_rg1.53
pddf_rg_errornull
guinier_rg1.53
guinier_rg_error0.004
pddf_dmax5.0
pddf_dmax_errornull
porod_volumenull
porod_volume_errornull
estimated_volumenull
estimated_volume_errornull
guinier_point_firstnull
guinier_point_lastnull
pddf_point_firstnull
pddf_point_lastnull
i0_calibration_standard_datanull
intensities_log_log_plotSASDAX6_datloglog_img.png
symmetrynull
last_modified2023-05-25T11:21:52.654103+02:00
bragg_peak[]
manifest.json:36 field values
Field pathRaw value
codeSASDAX6
statussuccess
started_at2026-08-11T15:17:47.960903+00:00
finished_at2026-08-11T15:17:57.429971+00:00
source_last_modified2023-05-25T11:21:52.654103+02:00
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View the complete summary.json source
{
  "code": "SASDAX6",
  "status": "Published",
  "type_of_curve": "Single concentration",
  "angular_unit": "1/nm",
  "project": {
    "title": "Structural bases for the interaction of frataxin with the central components of iron-sulphur cluster assembly.",
    "publication": {
      "title": "Structural bases for the interaction of frataxin with the central components of iron-sulphur cluster assembly.",
      "author_list": "Prischi F, Konarev PV, Iannuzzi C, Pastore C, Adinolfi S, Martin SR, Svergun DI, Pastore A",
      "journal": "Nat Commun",
      "doi": "10.1038/ncomms1097",
      "pmid": "20981023",
      "published_date": "2010 Oct 19"
    },
    "status": "released",
    "submitted_date": null,
    "released_date": "2015-07-07"
  },
  "pddf_data": "https://www.sasbdb.org/media/p_of_R_files/SASDAX6.out",
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        "resolution": 0.172
      },
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      "city": "Hamburg",
      "country": "Germany",
      "beamline_name": "EMBL X33",
      "beam_geometry": "0.60",
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    "sample": {
      "molecule": [
        {
          "long_name": "Protein CyaY",
          "short_name": null,
          "sequence": "MNDSEFHRLADQLWLTIEERLDDWDGDSDIDCEINGGVLTITFENGSKIIINRQEPLHQVWLATKQGGYHFDLKGDEWIC\r\nDRSGETFWDLLEQAATQQAGETVSFR",
          "organism": "Escherichia coli",
          "uniprot_code": "P27838",
          "uniprot_range_first": null,
          "uniprot_range_last": null,
          "oligomerization": "monomer",
          "molecular_type": "protein",
          "uniprot_sequence": "MNDSEFHRLADQLWLTIEERLDDWDGDSDIDCEINGGVLTITFENGSKIIINRQEPLHQV\nWLATKQGGYHFDLKGDEWICDRSGETFWDLLEQAATQQAGETVSFR",
          "mw": 12.231,
          "total_mw": 12.231,
          "number_molecules": 1,
          "complex_state": null,
          "deuteration": null,
          "molecule_source": "biological",
          "molecule_description": null
        }
      ],
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View the complete manifest.json source
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