Published assay observation

compound 5 (Collie 2025) and NSD3

PWWP11:ASSAY:0002

censored binding limit

Reported observation

SPR Kd counter-screen

> 30.0 µM

Target domain: UP:Q9BZ95:PWWP1
Assay construct: 263–398
Identity scope: assayed stereoisomeric mixture not single chemical identity

Evidence provenance

Primary source

PMID:40959233

Table 1 NSD3 (263–398) column

Verification scope: Phase 3 primary Table 1 and actual supporting PDF pages 7, 9 and 14; the inherited HTRF method remains explicitly not inspected

Assay and verification JSON

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	{"end": 398, "numbering": "canonical UniProt inclusive", "start": 263} verified	SV05B · Table S2, PDF page 7 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
tags	N-terminal 6His-TEV retained verified	SV05B · Table S2, PDF page 7 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
engineered mutations	not reported	SV05B · Table S2, PDF page 7 No substitutions are listed for these assay constructs. The separate crystallography construct for 9FOE lists seven mutations; those are not applied to assay records. This is not a sequence audit. Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
assay type	SPR Kd counter-screen verified	SV05A · Table 1 Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
temperature C	25 verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
buffer	10 mM HEPES; 150 mM NaCl; 3 mM EDTA; 0.1 mM TCEP; 0.05% (w/v) Tween-20; 0.3% (v/v) DMSO verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
pH	7.5 verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
DMSO percent	0.3 verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
units	µM verified	SV05A · Table 1 Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
measurement uncertainty	not applicable	SV05A · Table 1 Censored threshold is not a point estimate with an error bar. Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
uncertainty type	not applicable	SV05A · Table 1 NSD3 column Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
replicate definition	At least 3 independent replicates; numeric Kd summaries are mean and SD. The footnote supplies no separate exact n for individual negative/censored cells. verified	SV05A · Table 1 footnote a Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
measurement entity	PREP:COLLIE2025_5 verified	SV05A · Table 1 The reported result belongs to this experimental preparation and construct; structural CCD membership alone cannot transfer affinity. Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
concentration range	{"dilution_factor": 0.5, "maximum_uM": null, "points": 12} verified	SV05B · SPR method, PDF page 9 Twofold series; endpoint not stated in inspected protocol. A >30 uM reported Kd bound is not reinterpreted as the assay maximum. Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
incubation and flow	{"contact_s": 60, "cycles": "multi-cycle", "dissociation_s": 120, "flow_uL_per_min": 30} verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
fitting	Double reference and solvent correction; 1:1 kinetic fitting or steady-state Langmuir fitting; Insight Evaluation Software 4.0. Exact fit choice is not separately assigned to each table row. verified	SV05B · SPR method, PDF page 9 Inspected source scope Actual primary PDF obtained from official Europe PMC supplementary archive after direct ACS retrieval returned 403; Table S2, SPR, biochemical method and compound 11-13 chemistry visually checked
negative observation	{"endpoint": "Kd", "operator": "\u003e", "units": "uM", "value": 30} verified	SV05A · Table 1 NSD3 column Explicit censored affinity bound; preserve operator and do not treat as a point Kd. Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
identity assignment	assayed_stereoisomeric_mixture_not_single_chemical_identity verified	SV05A · Synthesis Results and Table 1 Existing curated exact identity, if present, is retained; this phase does not change chemical nodes. Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected

Structure → Component → Assay

Exact identity contact links

0 contact links

No exact-identity structural contact is linked to this observation. The preparation-level observation remains available.