Source was purified recombinant proteins and peptides in solution.
Source was purified Su(var)205 tagged protein and His3 tagged peptides in solution.
Results indicate that the chromo domains of the dimeric intact Su(var)205 protein act independently of each other.
Source was purified Su(var)205 tagged protein and His3 tagged peptide in solution.
V26M mutation in Su(var)205 protein completely abolishes interaction.
Source was purified recombinant Su(var)205 protein and His3 peptides in solution.
Source was purified recombinant Su(var)205 protein and His3 peptides in solution.
His3 protein residues flanking K9 (aa 6-8 and 10) contribute to chromodomain binding specificity.
Interaction in vitro; bait peptide produced commercially; prey produced as a recombinant fusion protein in bacterial system.
Bait expressed and purified from bacteria.
Prey endogenous to cells and purified as total nucleosomes.
A His3 N-terminal peptide bearing K9 trimethylation, but not unmodified peptide, competes with endogenous His3 for binding to Su(var)205 (HP1a).
Purified Su(var)205 was fractionated by PAGE, transferred to membrane, and tested for its ability to bind endogenous His3. Binding was assayed using anti-His3 antibodies.
Bait endogenous to cells and purified as nucleosomes.
Prey expressed and purified from bacteria.
Purified His3 was fractionated by PAGE, transferred to membrane, and tested for its ability to bind Su(var)205 (HP1a).
Bait expressed and purified from bacteria.
Prey endogenous to cells and purified as mononucleosomes.
Su(var)205-bound His3 is enriched for K9 trimethylation.
GST pull down.
Interaction in vitro; recombinant proteins produced in baculovirus and insect cell system.
Interaction detected with H3K9me2 and H3K9me3 not with H3K4me2 or H3K36me3
Source was purified Su(var)205 tagged protein and His3 tagged peptides in solution.
Analyzed Su(var)205 protein variants modifed at E52; results suggest that specific recognition of H3K9Me2 vs. H3K9Me3 is not necessary in this protein–protein interaction.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey peptide synthesized in vitro.
His3 peptides used in binding assays purchased from Diagenode, sequences unspecified.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey peptide chemically synthesized and labeled in vitro.
Interaction in vitro; bait produced and labeled in vitro; prey produced as a recombinant fusion protein in bacterial system.
Su(var)205 binds only the methylated His3 H3K9me3 peptide, not the unmethylated form.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.
Interaction in vitro; bait peptide chemically synthesized and labeled in vitro; prey produced as a recombinant fusion protein in baculovirus and Sf21 cell system.
For Su(var)205-His3K9me3, Kd = 1.0uM
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial or baculoviral system; prey peptide chemically synthesized and labeled in vitro.
K46A,E52D,D62F, coordinates relative to Su(var)205-PA, Kd changed from 1.2uM to 700nM.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey peptide synthesized in vitro.
K46A,E52D,D62F, coordinates relative to Su(var)205-PA
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey peptide synthesized in vitro.
K46A,E52D,D62F, coordinates relative to Su(var)205-PA
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey peptide synthesized in vitro.
