Published assay observation

compound 12 (Collie 2025) and NSD2

PWWP:ASSAY:0019

direct binding affinity

Reported observation

SPR Kd

= 1.47 µM ± 0.13

Reported uncertainty: SD

Target domain: UP:O96028:PWWP1
Assay construct: 208–368
Identity scope: paper label exact identity unresolved

Evidence provenance

Primary source

PMID:40959233

Table 1

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": 368, "numbering": "canonical UniProt inclusive", "start": 208} 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 Avi; C-terminal 6His; biotinylated 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 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	0.13 verified	SV05A · Table 1 Reported plus/minus magnitude, in the units of the assay result. Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
uncertainty type	SD verified	SV05A · Table 1 footnote a 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_12 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	not applicable	SV05A · Table 1 Inspected source scope Actual primary XML; full Results, Table 1 and footnotes inspected
identity assignment	{"absolute_sulfolane_configuration": null, "exact_ccd_id": null, "isomer_label": 1, "level": "source-labelled separated fraction", "reported_diastereomeric_ratio": "98.84:1.16", "retention_time_min": 7.1} verified	SV05B · Compounds 12/13, PDF page 14 Purified fractions are not assumed enantiopure. Retention time is not an absolute stereochemical assignment. 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

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.