6XCG V01
native annotation core contact
same exact chemical identity and native domain; construct equivalence not assumed
Published assay observation
UNC6934 and NSD2
PWWP:ASSAY:0006
Reported observation
NanoBRET PPI half-response
= 1.23 µM ± 0.25
Reported uncertainty; definition uncurated
- Target domain
- UP:O96028:PWWP1
- Assay construct
- Uncurated–Uncurated
- Identity scope
- exact single CCD identity
Evidence provenance
Primary source
PMID:34782742Results: cellular target engagement; Figure 4d
- Verification scope
- Phase 3 primary full-text Methods, Results and figure legends through official NCBI BioC; actual publisher supplement; assay-specific scope retained
Field-level provenance
Assay conditions and source verification
Missing information is not proof of absence. The v1.2 source overlay separates not reported in inspected material, not inspected, not transcribed and not applicable.
| Field | Value and source state | Source and interpretation |
|---|---|---|
| construct bounds | not reported | SV02A · Plasmids and NanoBRET PPI Methods Domain named but residue boundaries are not explicitly specified for the NanoLuc construct in inspected text; biochemical 208–368 is not copied onto a cellular construct. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| tags | C-terminal NanoLuc on NSD2-PWWP1; C-terminal HaloTag on histone H3.3 verified | SV02A · NanoBRET PPI MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| engineered mutations | WT assay; F266A tested separately verified | SV02A · Figure 4d/e and NanoBRET PPI MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| assay type | NanoBRET PPI half-response verified | SV02A · Results and assay-specific MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| temperature C | not reported | SV02A · Cell culture and NanoBRET PPI Methods Cell culture is at 37°C and 5% CO2; the 20 h assay and plate-reader temperatures are not explicitly restated. Keep culture conditions separate from a scalar assay temperature. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| buffer | DMEM/F12 without phenol red; 4% FBS; 100 U/mL penicillin; 100 ug/mL streptomycin verified | SV02A · NanoBRET PPI MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| pH | not inspected | Outside the source scope inspected for this field; no absence claim. |
| DMSO percent | not inspected | Outside the source scope inspected for this field; no absence claim. |
| units | µM verified | SV02A · Results and assay-specific MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| measurement uncertainty | 0.25 verified | SV02A · Results and assay-specific Methods Reported plus/minus magnitude, in the units of the assay result. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| uncertainty type | not reported | SV02A · Figure 4d; statistical analysis ±0.25 uM preserved; the inspected specific figure legend does not label its summary statistic. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| replicate definition | 3 independent replicates verified | SV02A · Figure 4dInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| measurement entity | PREP:UNC6934 verified | SV02A · Results and assay-specific Methods The reported result belongs to this experimental preparation and construct; structural CCD membership alone cannot transfer affinity. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| concentration range | not inspected | Outside the source scope inspected for this field; no absence claim. |
| incubation and flow | {"CO2_percent": 5, "cell_culture_C": 37, "compound_treatment_h": 20, "readout_after_substrate_min": "within 10"} verified | SV02A · NanoBRET PPI Methods and Cell cultureInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| fitting | Half-response inferred from NanoBRET signal; Results/Figure 4d call EC50, Discussion calls IC50 source conflict | SV02A · Results/Figure 4d and Discussion Retain neutral cellular half-response endpoint; do not equate this to direct-binding Kd. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| negative observation | not applicable | SV02A · Results and assay-specific MethodsInspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
| identity assignment | exact_single_CCD_identity verified | SV02A · Chemical series Results and existing CCD mapping No change to existing identity node or preparation ID. Inspected source scopePrimary author manuscript full text retrieved; relevant Methods, Results and figure legends inspected |
Structure → Component → Assay