[English] 日本語
Yorodumi
- EMDB-70552: Plunge frozen control Beta Galactosidase -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: EMDB / ID: EMD-70552
TitlePlunge frozen control Beta Galactosidase
Map dataUnsharpened Plunge frozen beta-galactosidase control
Sample
  • Complex: Standard plunge frozen Beta Galactosidase from E. coli
    • Protein or peptide: Beta Galactosidase
KeywordsComplex / Protein / HYDROLASE
Function / homology
Function and homology information


alkali metal ion binding / lactose catabolic process / beta-galactosidase complex / beta-galactosidase / beta-galactosidase activity / carbohydrate binding / magnesium ion binding / identical protein binding
Similarity search - Function
Glycoside hydrolase, family 2, beta-galactosidase / Beta galactosidase small chain/ domain 5 / Beta-galactosidase, domain 4 / Beta galactosidase small chain / Beta-galactosidase, domain 4 / Beta galactosidase small chain / : / Glycoside hydrolase, family 2, active site / Glycosyl hydrolases family 2 acid/base catalyst. / Glycoside hydrolase, family 2, conserved site ...Glycoside hydrolase, family 2, beta-galactosidase / Beta galactosidase small chain/ domain 5 / Beta-galactosidase, domain 4 / Beta galactosidase small chain / Beta-galactosidase, domain 4 / Beta galactosidase small chain / : / Glycoside hydrolase, family 2, active site / Glycosyl hydrolases family 2 acid/base catalyst. / Glycoside hydrolase, family 2, conserved site / Glycosyl hydrolases family 2 signature 1. / Glycoside hydrolase, family 2 / Glycosyl hydrolases family 2, sugar binding domain / Glycoside hydrolase family 2, catalytic domain / Glycosyl hydrolases family 2, sugar binding domain / Glycosyl hydrolases family 2, TIM barrel domain / Glycoside hydrolase, family 2, immunoglobulin-like beta-sandwich / Glycosyl hydrolases family 2 / Beta-Galactosidase/glucuronidase domain superfamily / Glycoside hydrolase-type carbohydrate-binding / Galactose mutarotase-like domain superfamily / Galactose-binding-like domain superfamily / Glycoside hydrolase superfamily / Immunoglobulin-like fold
Similarity search - Domain/homology
Biological speciesEscherichia coli (E. coli)
Methodsingle particle reconstruction / cryo EM / Resolution: 5.9 Å
AuthorsMertz KL / Jordahl D / Hemme CA / Probasco MD / Forbes DS / Ducos PL / Salome AZ / Quarmby ST / Grant T / Coon JJ
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35 GM118110 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)T32 GM135066 United States
National Science Foundation (NSF, United States)2137424 United States
CitationJournal: Mol Cell Proteomics / Year: 2025
Title: Laser-Induced Rehydration of Cryo-Landed Proteins Restores Native Structure.
Authors: Keaton L Mertz / Drew Jordahl / Colin A Hemme / Mitchell D Probasco / Dylan S Forbes / Peter L Ducos / Austin Z Salome / Michael S Westphall / Scott T Quarmby / Timothy Grant / Joshua J Coon /
Abstract: The use of native mass spectrometry (MS) to land biological molecules for subsequent cryogenic electron microscopy (cryoEM) imaging and three-dimensional reconstruction has gained momentum in recent ...The use of native mass spectrometry (MS) to land biological molecules for subsequent cryogenic electron microscopy (cryoEM) imaging and three-dimensional reconstruction has gained momentum in recent years as a means to overcome long-standing challenges posed by traditional cryoEM sample preparation. However, recent results obtained with this approach have been constrained by low resolution and the compaction of cryo-landed particles, likely due to dehydration during exposure to vacuum. Here, we describe a new sample preparation method that uses a laser integrated into a cryogenic soft-landing apparatus to liquefy precisely deposited amorphous ice, rehydrating particles, and restoring their solution structure prior to rapid revitrification via the thermal mass of the grid. With this technique, we demonstrate the reconstruction of cryo-landed, rehydrated, and revitrified β-galactosidase that is comparable in resolution to that achieved with plunge freezing. Furthermore, these particles are not compacted, matching the known structure and conformation obtained with traditionally plunge-frozen particles. These results establish the viability of coupling native MS with cryoEM for high-resolution structural determination without the limitations imposed by conventional sample preparation, and they open a path to solving previously inaccessible molecules and to integrating MS capabilities such as gas-phase purification to complex samples such as cell lysates.
History
DepositionMay 8, 2025-
Header (metadata) releaseJun 11, 2025-
Map releaseJun 11, 2025-
UpdateJun 11, 2025-
Current statusJun 11, 2025Processing site: RCSB / Status: Released

-
Structure visualization

Supplemental images

emd_70552.png

Downloads & links

-
Map

FileDownload / File: emd_70552.map.gz / Format: CCP4 / Size: 437.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationUnsharpened Plunge frozen beta-galactosidase control
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.2 Å/pix.
x 486 pix.
= 583.2 Å		1.2 Å/pix.
x 486 pix.
= 583.2 Å		1.2 Å/pix.
x 486 pix.
= 583.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.2 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 2.95
Minimum - Maximum-5.9735365 - 12.923126
Average (Standard dev.)-0.007785428 (±0.5820123)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions486486486
Spacing486486486
CellA=B=C: 583.2 Å
α=β=γ: 90.0 °

-
Supplemental data

-
Half map: Plunge frozen beta-galactosidase control half map 2

Fileemd_70552_half_map_1.map
AnnotationPlunge frozen beta-galactosidase control half map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Half map: Plunge frozen beta-galactosidase control half map 1

Fileemd_70552_half_map_2.map
AnnotationPlunge frozen beta-galactosidase control half map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Sample components

-
Entire : Standard plunge frozen Beta Galactosidase from E. coli

EntireName: Standard plunge frozen Beta Galactosidase from E. coli
Components
  • Complex: Standard plunge frozen Beta Galactosidase from E. coli
    • Protein or peptide: Beta Galactosidase

-
Supramolecule #1: Standard plunge frozen Beta Galactosidase from E. coli

SupramoleculeName: Standard plunge frozen Beta Galactosidase from E. coli
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Escherichia coli (E. coli)
Molecular weightTheoretical: 465 KDa

-
Macromolecule #1: Beta Galactosidase

MacromoleculeName: Beta Galactosidase / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Escherichia coli (E. coli)
SequenceString: MTMITDSLAV VLQRRDWENP GVTQLNRLAA HPPFASWRNS EEARTDRPSQ QLRSLNGEWR FAWFPAPEA VPESWLECDL PEADTVVVPS NWQMHGYDAP IYTNVTYPIT VNPPFVPTEN P TGCYSLTF NVDESWLQEG QTRIIFDGVN SAFHLWCNGR WVGYGQDSRL ...String:
MTMITDSLAV VLQRRDWENP GVTQLNRLAA HPPFASWRNS EEARTDRPSQ QLRSLNGEWR FAWFPAPEA VPESWLECDL PEADTVVVPS NWQMHGYDAP IYTNVTYPIT VNPPFVPTEN P TGCYSLTF NVDESWLQEG QTRIIFDGVN SAFHLWCNGR WVGYGQDSRL PSEFDLSAFL RA GENRLAV MVLRWSDGSY LEDQDMWRMS GIFRDVSLLH KPTTQISDFH VATRFNDDFS RAV LEAEVQ MCGELRDYLR VTVSLWQGET QVASGTAPFG GEIIDERGGY ADRVTLRLNV ENPK LWSAE IPNLYRAVVE LHTADGTLIE AEACDVGFRE VRIENGLLLL NGKPLLIRGV NRHEH HPLH GQVMDEQTMV QDILLMKQNN FNAVRCSHYP NHPLWYTLCD RYGLYVVDEA NIETHG MVP MNRLTDDPRW LPAMSERVTR MVQRDRNHPS VIIWSLGNES GHGANHDALY RWIKSVD PS RPVQYEGGGA DTTATDIICP MYARVDEDQP FPAVPKWSIK KWLSLPGETR PLILCEYA H AMGNSLGGFA KYWQAFRQYP RLQGGFVWDW VDQSLIKYDE NGNPWSAYGG DFGDTPNDR QFCMNGLVFA DRTPHPALTE AKHQQQFFQF RLSGQTIEVT SEYLFRHSDN ELLHWMVALD GKPLASGEV PLDVAPQGKQ LIELPELPQP ESAGQLWLTV RVVQPNATAW SEAGHISAWQ Q WRLAENLS VTLPAASHAI PHLTTSEMDF CIELGNKRWQ FNRQSGFLSQ MWIGDKKQLL TP LRDQFTR APLDNDIGVS EATRIDPNAW VERWKAAGHY QAEAALLQCT ADTLADAVLI TTA HAWQHQ GKTLFISRKT YRIDGSGQMA ITVDVEVASD TPHPARIGLN CQLAQVAERV NWLG LGPQE NYPDRLTAAC FDRWDLPLSD MYTPYVFPSE NGLRCGTREL NYGPHQWRGD FQFNI SRYS QQQLMETSHR HLLHAEEGTW LNIDGFHMGI GGDDSWSPSV SAEFQLSAGR YHYQLV WCQ K

UniProtKB: Beta-galactosidase

-
Experimental details

-
Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

Concentration1 mg/mL
BufferpH: 7.6 / Component - Concentration: 100.0 mM / Component - Formula: NH4CH3CO2 / Component - Name: Ammonium Acetate
GridModel: UltrAuFoil / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: LACEY / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec.
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV

-
Electron microscopy

MicroscopeTFS GLACIOS
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Average electron dose: 36.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm
Sample stageCooling holder cryogen: NITROGEN

+
Image processing

CTF correctionSoftware - Name: cisTEM / Type: NONE
Startup modelType of model: NONE / Details: Ab initio
Final reconstructionApplied symmetry - Point group: D2 (2x2 fold dihedral) / Resolution.type: BY AUTHOR / Resolution: 5.9 Å / Resolution method: OTHER / Software - Name: cisTEM / Details: Map-to-model FSC, PDB 6CVM / Number images used: 5684
Initial angle assignmentType: PROJECTION MATCHING / Software - Name: cisTEM
Final angle assignmentType: PROJECTION MATCHING / Software - Name: cisTEM
Final 3D classificationNumber classes: 30 / Software - Name: cisTEM
FSC plot (resolution estimation)

+
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