SASDCS7: TET12(1.10)SN-f9

Basic information

• Entry: Database: SASBDB / ID: SASDCS7

Sample

TET12(1.10)SN-f9

• TET12(1.10)SN-f9 (protein), synthetic construct

• Biological species: synthetic construct (others)
• Citation: Journal: Nat Biotechnol / Year: 2017; Title: Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo.; Authors: Ajasja Ljubetič / Fabio Lapenta / Helena Gradišar / Igor Drobnak / Jana Aupič / Žiga Strmšek / Duško Lainšček / Iva Hafner-Bratkovič / Andreja Majerle / Nuša Krivec / Mojca Benčina / Tomaž Pisanski / Tanja Ćirković Veličković / Adam Round / José María Carazo / Roberto Melero / Roman Jerala / ; Abstract: Polypeptides and polynucleotides are natural programmable biopolymers that can self-assemble into complex tertiary structures. We describe a system analogous to designed DNA nanostructures in which protein coiled-coil (CC) dimers serve as building blocks for modular de novo design of polyhedral protein cages that efficiently self-assemble in vitro and in vivo. We produced and characterized >20 single-chain protein cages in three shapes-tetrahedron, four-sided pyramid, and triangular prism-with the largest containing >700 amino-acid residues and measuring 11 nm in diameter. Their stability and folding kinetics were similar to those of natural proteins. Solution small-angle X-ray scattering (SAXS), electron microscopy (EM), and biophysical analysis confirmed agreement of the expressed structures with the designs. We also demonstrated self-assembly of a tetrahedral structure in bacteria, mammalian cells, and mice without evidence of inflammation. A semi-automated computational design platform and a toolbox of CC building modules are provided to enable the design of protein cages in any polyhedral shape.

Contact author

• Jana Aupic (National Institute of Chemistry, Slovenia)

Models

• Model #1577: ; Type: atomic / Software: (9.16) / Radius of dummy atoms: 1.90 A; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: TET12(1.10)SN-f9 / Specimen concentration: 1.60-6.00
• Buffer: Name: 20 mM Tris 150 mM NaCl 10% glycerol / pH: 7.5

Entity #803

Type: protein / Description: TET12(1.10)SN-f9 / Formula weight: 56.581 / Num. of mol.: 1 / Source: synthetic construct
Sequence:

MLEEELKQLE EELQAIEEQL AQLQWKAQAR KEKLAQLKEK LSGSGPGSGS PEDEIQQLEE EISQLEQKNS ELKEKNQELK YGSGSGPGSG DIEQELERAK ESIRRLEQEV NQERSRMQYL QTLLEKSGSG PGSGQLEDKV EELLSKNYHL ENEVERLKKL VGSGSGPGSG LEEELKQLEE ELQAIEEQLA QLQWKAQARK EKLAQLKEKL SGSGPGSGSP EDEIQQLEEK NSQLKQEISQ LEEKNQELKY GSGSGPGSGQ LEDKVEELLS KNYHLENEVE RLKKLVGSGS GPGSGSPEDK ISQLKEKIQQ LKQENQQLEE ENSQLEYGSG SGPGSGSPED ENSQLEEKIS QLKQKNSELK EEIQQLEYGS GSGPGSGSPE DKISELKEEN QQLEQKIQQL KEENSQLEYG SGSGPGSGDI EQELERAKES IRRLEQEVNQ ERSRMQYLQT LLEKSGSGPG SGSPEDKNSE LKEEIQQLEE ENQQLEEKIS ELKYGLEHHH HHHHH

Experimental information

• Beam: Instrument name: PETRA III P12 / City: Hamburg / 国: Germany / Shape: Point / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.124 Å / Dist. spec. to detc.: 2 mm
• Detector: Name: Pilatus 1M-W / Pixsize x: 0.172 mm

Scan

Title: TET12(1.10)SN-f9 / Measurement date: Jun 10, 2016 / Storage temperature: 10 °C / Cell temperature: 20 °C / Exposure time: 0.045 sec. / Number of frames: 20 / Unit: 1/nm /

	Min	Max
Q	0.2526	6.7156

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 291 /

	Min	Max
Q	0.252581	1.83291
P(R) point	1	291
R	0	12.9

Result

Type of curve: single_conc

	Experimental	Standard	Standard error	Porod
MW	37 kDa	37 kDa	2	90 kDa
Volume	-	-	-	153 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	5027	20	5021	60
Radius of gyration, Rg	3.95 nm	0.01	3.87 nm	0.05

	Min	Max	Error
D	-	12.9	2
Guinier point	3	16	-