6SH9

EngBF DARPin Fusion 4b D12

Summary for 6SH9

• Entry DOI: 10.2210/pdb6sh9/pdb
• Related: 6QEP
• Descriptor: Endo-alpha-N-acetylgalactosaminidase,DARPin 4b D12, Envelope glycoprotein gp160, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (6 entities in total)
• Functional Keywords: crystallization chaperone, protein fusion, darpin, chaperone, hydrolase
• Biological source: Bifidobacterium longum subsp. longum JCM 1217; More
• Total number of polymer chains: 2
• Total formula weight: 150064.78

Authors

Ernst, P.,Pluckthun, A.,Mittl, P.R.E. (deposition date: 2019-08-06, release date: 2019-11-06, Last modification date: 2024-01-24)

Primary citation

Ernst, P.,Pluckthun, A.,Mittl, P.R.E.
Structural analysis of biological targets by host:guest crystal lattice engineering.
Sci Rep, 9:15199-15199, 2019

PubMed Abstract: To overcome the laborious identification of crystallisation conditions for protein X-ray crystallography, we developed a method where the examined protein is immobilised as a guest molecule in a universal host lattice. We applied crystal engineering to create a generic crystalline host lattice under reproducible, predefined conditions and analysed the structures of target guest molecules of different size, namely two 15-mer peptides and green fluorescent protein (sfGFP). A fusion protein with an N-terminal endo-α-N-acetylgalactosaminidase (EngBF) domain and a C-terminal designed ankyrin repeat protein (DARPin) domain establishes the crystal lattice. The target is recruited into the host lattice, always in the same crystal form, through binding to the DARPin. The target structures can be determined rapidly from difference Fourier maps, whose quality depends on the size of the target and the orientation of the DARPin.

PubMed: 31645583
DOI: 10.1038/s41598-019-51017-y

Experimental method

X-RAY DIFFRACTION (2.4 Å)