Summary information and primary citation
- PDB-id
-
8tiu;
DSSR-derived features in text and
JSON formats
- Class
- DNA binding protein-DNA
- Method
- X-ray (3.9 Å)
- Summary
- Isoreticular, interpenetrating co-crystal of
replication initiator protein repe54 and symmetrical
expanded duplex (31mer) containing the cognate repe54
sequence and an additional g-c rich sequence with 2 sticky
base overhangs and 3' terminal phosphates.
- Reference
-
Orun AR, Slaughter CK, Shields ET, Vajapayajula A, Jones
S, Shrestha R, Snow CD (2024): "Tuning
Chemical DNA Ligation within DNA Crystals and Protein-DNA
Cocrystals." Acs Nanosci Au,
4, 338-348. doi: 10.1021/acsnanoscienceau.4c00013.
- Abstract
- Biomolecular crystals can serve as materials for a
plethora of applications including precise guest
entrapment. However, as grown, biomolecular crystals are
fragile in solutions other than their growth conditions.
For crystals to achieve their full potential as hosts for
other molecules, crystals can be made stronger with
bioconjugation. Building on our previous work using
carbodiimide
1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC) for
chemical ligation, here, we investigate DNA junction
architecture through sticky base overhang lengths and the
role of scaffold proteins in cross-linking within two
classes of biomolecular crystals: cocrystals of DNA-binding
proteins and pure DNA crystals. Both crystal classes
contain DNA junctions where DNA strands stack up
end-to-end. Ligation yields were studied as a function of
sticky base overhang length and terminal phosphorylation
status. The best ligation performance for both crystal
classes was achieved with longer sticky overhangs and
terminal 3'phosphates. Notably, EDC chemical ligation was
achieved in crystals with pore sizes too small for
intracrystal transport of ligase enzyme. Postassembly
cross-linking produced dramatic stability improvements for
both DNA crystals and cocrystals in water and blood serum.
The results presented may help crystals containing DNA
achieve broader application utility, including as
structural biology scaffolds.
