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TitleA 3.8 Å resolution cryo-EM structure of a small protein bound to an imaging scaffold.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 1864, Year 2019
Publish dateApr 23, 2019
AuthorsYuxi Liu / Duc T Huynh / Todd O Yeates /
PubMed AbstractProteins smaller than about 50 kDa are currently too small to be imaged at high resolution by cryo-electron microscopy (cryo-EM), leaving most protein molecules in the cell beyond the reach of this ...Proteins smaller than about 50 kDa are currently too small to be imaged at high resolution by cryo-electron microscopy (cryo-EM), leaving most protein molecules in the cell beyond the reach of this powerful structural technique. Here we use a designed protein scaffold to bind and symmetrically display 12 copies of a small 26 kDa protein, green fluorescent protein (GFP). We show that the bound cargo protein is held rigidly enough to visualize it at a resolution of 3.8 Å by cryo-EM, where specific structural features of the protein are visible. The designed scaffold is modular and can be modified through modest changes in its amino acid sequence to bind and display diverse proteins for imaging, thus providing a general method to break through the lower size limitation in cryo-EM.
External linksNat Commun / PubMed:31015551 / PubMed Central
MethodsEM (single particle)
Resolution2.9 - 3.5 Å
Structure data

EMDB-9373, PDB-6nht:
Single particle reconstruction of the symmetric core an engineered protein scaffold
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-9374, PDB-6nhv:
Single particle reconstruction of DARPin and its bound GFP on a symmetric scaffold
Method: EM (single particle) / Resolution: 3.5 Å

Source
  • Pseudomonas aeruginosa PAO1 (bacteria)
  • Pyrococcus horikoshii OT3 (archaea)
  • pyrococcus horikoshii (strain atcc 700860 / dsm 12428 / jcm 9974 / nbrc 100139 / ot-3) (archaea)
  • pseudomonas aeruginosa (strain atcc 15692 / dsm 22644 / cip 104116 / jcm 14847 / lmg 12228 / 1c / prs 101 / pao1) (bacteria)
  • aequorea victoria (jellyfish)
KeywordsBIOSYNTHETIC PROTEIN / protein engineering / symmetric scaffold / small protein cryo-EM / display platform

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