[English] 日本語
Yorodumi
- PDB-5uwh: Crystal Structure of Paxillin NES Peptide in complex with CRM1-Ra... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: PDB / ID: 5uwh
TitleCrystal Structure of Paxillin NES Peptide in complex with CRM1-Ran-RanBP1
Components
  • Exportin-1
  • GTP-binding nuclear protein Ran
  • Paxillin
  • Ran-specific GTPase-activating protein 1
KeywordsPROTEIN TRANSPORT / HEAT repeat / NES / nuclear export / Karyopherin
Function / homology
Function and homology information


Regulation of cytoskeletal remodeling and cell spreading by IPP complex components / Localization of the PINCH-ILK-PARVIN complex to focal adhesions / Regulation of MITF-M-dependent genes involved in extracellular matrix, focal adhesion and epithelial-to-mesenchymal transition / tRNA re-export from nucleus / Transport of Mature mRNA derived from an Intron-Containing Transcript / pre-miRNA export from nucleus / RNA nuclear export complex / snRNA import into nucleus / Regulation of HSF1-mediated heat shock response / nuclear export signal receptor activity ...Regulation of cytoskeletal remodeling and cell spreading by IPP complex components / Localization of the PINCH-ILK-PARVIN complex to focal adhesions / Regulation of MITF-M-dependent genes involved in extracellular matrix, focal adhesion and epithelial-to-mesenchymal transition / tRNA re-export from nucleus / Transport of Mature mRNA derived from an Intron-Containing Transcript / pre-miRNA export from nucleus / RNA nuclear export complex / snRNA import into nucleus / Regulation of HSF1-mediated heat shock response / nuclear export signal receptor activity / Transcriptional and post-translational regulation of MITF-M expression and activity / manchette / cellular response to mineralocorticoid stimulus / neuropilin binding / vinculin binding / Regulation of cholesterol biosynthesis by SREBP (SREBF) / tRNA export from nucleus / importin-alpha family protein binding / SUMOylation of SUMOylation proteins / protein localization to kinetochore / spindle pole body / Rev-mediated nuclear export of HIV RNA / signal complex assembly / Nuclear import of Rev protein / U4 snRNA binding / nuclear export / SUMOylation of RNA binding proteins / protein localization to nucleolus / NEP/NS2 Interacts with the Cellular Export Machinery / RNA export from nucleus / tRNA processing in the nucleus / GTP metabolic process / Postmitotic nuclear pore complex (NPC) reformation / SUMOylation of chromatin organization proteins / nuclear import signal receptor activity / microtubule associated complex / growth hormone receptor signaling pathway / MicroRNA (miRNA) biogenesis / DNA metabolic process / MAPK6/MAPK4 signaling / dynein intermediate chain binding / mitotic sister chromatid segregation / ribosomal large subunit export from nucleus / U5 snRNA binding / spermatid development / Smooth Muscle Contraction / viral process / GAB1 signalosome / U2 snRNA binding / U6 snRNA binding / positive regulation of protein binding / endothelial cell migration / sperm flagellum / nuclear pore / mRNA export from nucleus / U1 snRNA binding / PTK6 Regulates RHO GTPases, RAS GTPase and MAP kinases / ribosomal subunit export from nucleus / stress fiber / ribosomal small subunit export from nucleus / positive regulation of stress fiber assembly / centriole / substrate adhesion-dependent cell spreading / transforming growth factor beta receptor signaling pathway / GTPase activator activity / protein export from nucleus / mitotic spindle organization / cellular response to reactive oxygen species / male germ cell nucleus / hippocampus development / Transcriptional regulation by small RNAs / G1/S transition of mitotic cell cycle / recycling endosome / beta-catenin binding / kinetochore / positive regulation of protein import into nucleus / small GTPase binding / VEGFA-VEGFR2 Pathway / protein import into nucleus / cell-cell junction / GDP binding / melanosome / nuclear envelope / cell migration / mitotic cell cycle / lamellipodium / G protein activity / actin cytoskeleton organization / ubiquitin-dependent protein catabolic process / midbody / cell cortex / protein phosphatase binding / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / cell adhesion / cadherin binding / protein heterodimerization activity / protein domain specific binding / cell division / focal adhesion / GTPase activity
Similarity search - Function
Paxillin / : / : / Paxillin family / Ran-specific GTPase-activating protein 1, Ran-binding domain / Exportin-1, repeat 3 / Chromosome region maintenance repeat / Exportin-1, repeat 2 / Chromosome region maintenance or exportin repeat / CRM1 / Exportin repeat 2 ...Paxillin / : / : / Paxillin family / Ran-specific GTPase-activating protein 1, Ran-binding domain / Exportin-1, repeat 3 / Chromosome region maintenance repeat / Exportin-1, repeat 2 / Chromosome region maintenance or exportin repeat / CRM1 / Exportin repeat 2 / CRM1 / Exportin repeat 3 / CRM1 C terminal / Exportin-1, C-terminal / CRM1 C terminal / Exportin-1/5 / Exportin-1/Importin-beta-like / Exportin 1-like protein / Ran binding protein RanBP1-like / Ran binding domain / RanBP1 domain / Ran binding domain type 1 profile. / Ran-binding domain / LIM zinc-binding domain signature. / Ran GTPase / Small GTPase Ran-type domain profile. / LIM domain / Importin-beta N-terminal domain / Importin-beta N-terminal domain / Importin-beta N-terminal domain profile. / Zinc-binding domain present in Lin-11, Isl-1, Mec-3. / Zinc finger, LIM-type / LIM domain profile. / Importin-beta, N-terminal domain / Pleckstrin-homology domain (PH domain)/Phosphotyrosine-binding domain (PTB) / PH-domain like / Ran (Ras-related nuclear proteins) /TC4 subfamily of small GTPases / Rho (Ras homology) subfamily of Ras-like small GTPases / Ras subfamily of RAS small GTPases / Small GTPase / Ras family / Rab subfamily of small GTPases / Armadillo-like helical / Small GTP-binding protein domain / PH-like domain superfamily / Armadillo-type fold / Roll / P-loop containing nucleotide triphosphate hydrolases / Rossmann fold / P-loop containing nucleoside triphosphate hydrolase / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER / Exportin-1 / Ran-specific GTPase-activating protein 1 / Paxillin / GTP-binding nuclear protein Ran
Similarity search - Component
Biological speciesHomo sapiens (human)
Saccharomyces cerevisiae (brewer's yeast)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.26 Å
AuthorsFung, H.Y.J. / Chook, Y.M.
Funding support United States, Hong Kong, 7items
OrganizationGrant numberCountry
Howard Hughes Medical Institute (HHMI)International Student Research Fellowship United States
Croucher FoundationPredoc Research Scholarship Hong Kong
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM069909 United States
Welch FoundationI-1532 United States
Leukemia & Lymphoma SocietyScholar Award United States
University of Texas Southwestern Medical CenterEndowed Scholars Program United States
Cancer Prevention and Research Institute of Texas (CPRIT)RP120352, RP150053 United States
CitationJournal: Elife / Year: 2017
Title: Nuclear export receptor CRM1 recognizes diverse conformations in nuclear export signals.
Authors: Fung, H.Y. / Fu, S.C. / Chook, Y.M.
History
DepositionFeb 21, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 22, 2017Provider: repository / Type: Initial release
Revision 1.1Sep 13, 2017Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.2Sep 27, 2017Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.3Apr 24, 2019Group: Author supporting evidence / Data collection / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.4Nov 20, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.5Oct 4, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

-
Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

-
Assembly

Deposited unit
A: GTP-binding nuclear protein Ran
B: Ran-specific GTPase-activating protein 1
C: Exportin-1
D: Paxillin
hetero molecules


Theoretical massNumber of molelcules
Total (without water)164,0609
Polymers163,2374
Non-polymers8235
Water8,917495
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area11460 Å2
ΔGint-60 kcal/mol
Surface area57760 Å2
MethodPISA
Unit cell
Length a, b, c (Å)106.984, 106.984, 304.880
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number96
Space group name H-MP43212
Components on special symmetry positions
IDModelComponents
11C-1554-

HOH

-
Components

-
Protein , 3 types, 3 molecules ABC

#1: Protein GTP-binding nuclear protein Ran / Androgen receptor-associated protein 24 / GTPase Ran / Ras-like protein TC4 / Ras-related nuclear protein


Mass: 26758.564 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: RAN, ARA24, OK/SW-cl.81 / Plasmid: pET15 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P62826
#2: Protein Ran-specific GTPase-activating protein 1 / Chromosome stability protein 20 / Perinuclear array-localized protein / Ran-binding protein 1 / RANBP1


Mass: 16521.867 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast)
Gene: YRB1, CST20, HTN1, SFO1, YDR002W, YD8119.08 / Plasmid: pGEX-4T3-TEV / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P41920
#3: Protein Exportin-1 / Chromosome region maintenance protein 1 / Karyopherin-124


Mass: 117458.781 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast)
Gene: CRM1, KAP124, XPO1, YGR218W, G8514 / Plasmid: pGEX-4T3-TEV / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P30822

-
Protein/peptide , 1 types, 1 molecules D

#4: Protein/peptide Paxillin


Mass: 2497.800 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: PXN / Plasmid: pMAL-TEV / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P49023*PLUS

-
Non-polymers , 4 types, 500 molecules

#5: Chemical ChemComp-GNP / PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER


Mass: 522.196 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Formula: C10H17N6O13P3
Comment: GppNHp, GMPPNP, energy-carrying molecule analogue*YM
#6: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Formula: Mg
#7: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C3H8O3
#8: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 495 / Source method: isolated from a natural source / Formula: H2O

-
Experimental details

-
Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

-
Sample preparation

CrystalDensity Matthews: 2.66 Å3/Da / Density % sol: 53.83 %
Crystal growTemperature: 295 K / Method: vapor diffusion, hanging drop / pH: 6.4
Details: 16% PEG3350, 100 mM Bis-Tris, pH 6.4, 200 mM ammonium nitrate, 20 mM HCl

-
Data collection

DiffractionMean temperature: 93 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.9795 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Jul 16, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 2.26→50 Å / Num. obs: 77349 / % possible obs: 91.8 % / Redundancy: 7.3 % / Biso Wilson estimate: 29.8919033835 Å2 / Rmerge(I) obs: 0.081 / Rpim(I) all: 0.031 / Net I/σ(I): 22.8
Reflection shellResolution: 2.26→2.3 Å / Redundancy: 7 % / Num. unique all: 3826 / Num. unique obs: 3826 / CC1/2: 0.621 / Rpim(I) all: 0.461 / % possible all: 92.7

-
Processing

Software
NameVersionClassification
PHENIX(1.11.1_2575: ???)refinement
HKL-3000data collection
HKL-3000data scaling
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 4HB2

4hb2


Resolution: 2.26→47.266 Å / SU ML: 0.26 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 21.72
RfactorNum. reflection% reflection
Rfree0.2182 2000 2.67 %
Rwork0.1821 --
obs0.1831 74848 88.92 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Refinement stepCycle: LAST / Resolution: 2.26→47.266 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms10903 0 18 495 11416
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00311347
X-RAY DIFFRACTIONf_angle_d0.55415387
X-RAY DIFFRACTIONf_dihedral_angle_d14.786897
X-RAY DIFFRACTIONf_chiral_restr0.0391752
X-RAY DIFFRACTIONf_plane_restr0.0031960
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.2564-2.31280.29851140.24744149X-RAY DIFFRACTION72
2.3128-2.37530.26991320.22674798X-RAY DIFFRACTION83
2.3753-2.44520.29791390.22215066X-RAY DIFFRACTION88
2.4452-2.52420.24461430.2145203X-RAY DIFFRACTION90
2.5242-2.61440.29651450.19955288X-RAY DIFFRACTION92
2.6144-2.7190.26881460.19465329X-RAY DIFFRACTION92
2.719-2.84280.22711470.19335377X-RAY DIFFRACTION92
2.8428-2.99260.23731470.19915328X-RAY DIFFRACTION92
2.9926-3.18010.25471470.18965362X-RAY DIFFRACTION92
3.1801-3.42550.21751480.18475368X-RAY DIFFRACTION92
3.4255-3.77010.2031470.17535382X-RAY DIFFRACTION92
3.7701-4.31540.18781480.15455403X-RAY DIFFRACTION91
4.3154-5.43560.17871480.14665392X-RAY DIFFRACTION90
5.4356-47.27660.18371490.18025403X-RAY DIFFRACTION86
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL11 (°2)L12 (°2)L13 (°2)L22 (°2)L23 (°2)L33 (°2)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T11 (Å2)T12 (Å2)T13 (Å2)T22 (Å2)T23 (Å2)T33 (Å2)Origin x (Å)Origin y (Å)Origin z (Å)
11.0012-0.8415-0.21992.42860.21612.25480.0310.3186-0.1133-0.43960.1313-0.6772-0.25070.3882-0.15860.2557-0.07930.1340.4309-0.14790.43517.577545.817930.0291
21.0104-1.4228-0.36454.590.6062.34780.25480.06170.12530.06030.1291-1.0588-0.55510.5139-0.40090.2939-0.11450.10620.3867-0.17540.620911.919253.205638.3094
32.0974-0.53990.00220.8015-0.56730.48720.09690.1092-0.28640.01210.0562-0.62570.13610.2842-0.03130.1859-0.0022-0.02390.3817-0.16930.58248.282337.360537.3937
41.6905-1.10390.61313.3978-1.84644.39130.10110.0479-0.2717-0.10290.0139-0.17760.37720.098-0.1070.13230.0287-0.02080.2697-0.09940.3239-1.404234.238638.8875
53.0919-0.29170.79633.4107-0.56174.5606-0.1513-0.3355-0.06820.49690.1037-0.1988-0.13350.1003-0.03660.19670.0252-0.0230.3122-0.11450.33271.679143.58644.1318
61.75930.734-0.61622.4483-0.56364.45540.10120.19580.0442-0.28-0.03050.29830.0262-0.7598-0.20480.18790.0398-0.06830.3375-0.08210.2583-11.160641.642334.351
71.79790.07440.06932.4497-0.05922.36220.1173-0.01750.121-0.12590.036-0.2054-0.33110.0146-0.10290.19210.00040.01760.2865-0.08010.2457-1.715352.162138.4428
82.09871.4406-2.65381.0603-1.88363.40540.5278-0.05010.61070.00520.15890.1508-1.1231.2709-0.42810.5859-0.29580.14380.7466-0.13350.602610.268868.011934.0755
97.60032.4712-0.52453.0946-1.57882.22220.28930.20360.4973-0.19590.3288-0.2164-0.5890.8811-0.36570.5696-0.21170.25240.7594-0.24870.60539.175559.51147.8664
100.0825-0.0416-0.32370.00790.18082.13120.0947-0.16520.11560.03890.1484-0.2498-0.73240.7599-0.22750.7883-0.17890.29920.6077-0.05150.664.152673.254930.4305
110.84080.95680.68031.36910.51870.87880.02850.16240.1648-0.1450.1240.088-0.77590.7315-0.04260.7511-0.25020.32020.5527-0.04840.55628.127669.49416.8457
120.9726-0.00070.09731.62270.57170.5368-0.0586-0.1914-0.05910.9070.3338-0.97080.49770.3697-0.0920.25670.1734-0.36930.4525-0.16030.718210.130825.052153.298
132.224-0.5510.81381.8918-0.13931.93760.05760.0116-0.03410.15210.0155-0.02020.13630.0088-0.08110.2178-0.02040.00650.3017-0.08370.1317-21.595145.929754.6673
140.7534-0.1735-0.65860.73250.54091.99020.21380.05690.2286-0.2363-0.0058-0.125-0.5314-0.1679-0.11290.35270.07080.06420.32070.040.2536-28.228258.893513.7361
150.9401-0.78180.8171.1656-1.3632.15570.14110.1015-0.2786-0.12490.06870.13290.08550.0384-0.14150.22130.01570.02250.2654-0.08940.2994-8.641717.520714.745
169.09382.0588-3.12224.4197-1.0614.20760.18-0.02480.32190.14-0.04960.2854-0.3259-0.042-0.10441.07240.25880.30120.56690.0150.8026-39.333376.330728.696
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 9 through 44 )
2X-RAY DIFFRACTION2chain 'A' and (resid 45 through 66 )
3X-RAY DIFFRACTION3chain 'A' and (resid 67 through 91 )
4X-RAY DIFFRACTION4chain 'A' and (resid 92 through 111 )
5X-RAY DIFFRACTION5chain 'A' and (resid 112 through 122 )
6X-RAY DIFFRACTION6chain 'A' and (resid 123 through 142 )
7X-RAY DIFFRACTION7chain 'A' and (resid 143 through 169 )
8X-RAY DIFFRACTION8chain 'A' and (resid 170 through 193 )
9X-RAY DIFFRACTION9chain 'A' and (resid 194 through 216 )
10X-RAY DIFFRACTION10chain 'B' and (resid 64 through 97 )
11X-RAY DIFFRACTION11chain 'B' and (resid 98 through 198 )
12X-RAY DIFFRACTION12chain 'C' and (resid -1 through 245 )
13X-RAY DIFFRACTION13chain 'C' and (resid 246 through 478 )
14X-RAY DIFFRACTION14chain 'C' and (resid 479 through 808 )
15X-RAY DIFFRACTION15chain 'C' and (resid 809 through 1053 )
16X-RAY DIFFRACTION16chain 'D' and (resid 265 through 278 )

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242)：Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more