Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

Publication

A foundational atlas of autism protein interactions reveals molecular convergence

Wang B, Vartak R, Zaltsman Y, Naing ZZC, Hennick KM, Polacco BJ, Bashir A, Eckhardt M, Bouhaddou M, Xu J, Sun N, Lasser M, Zhou Y, Guiley KZ, Chan U, Kaye JA, Khare P, Drake S, Drury V, Burke DF, Gonzalez S, Alkhairy S, Morris M, Baum T, Krasnoff R, Wang S, Pham P, Arbalaez J, Pratt D, Chag S, Rolland T, Bourgeron T, Finkbeiner S, Bandyopadhay S, Ideker T, Beltrao P, Willsey HR, Obernier K, Nowakowski TJ, Huttenhain R, State MW, Willsey AJ, Krogan NJ

Translating high-confidence (hc) autism spectrum disorder (ASD) genes into viable treatment targets remains elusive. We constructed a foundational protein-protein interaction (PPI) network in HEK293T cells involving 100 hcASD risk genes, revealing over 1,800 PPIs (87% novel). Interactors, expressed in the human brain and enriched for ASD but not schizophrenia genetic risk, converged on protein complexes involved in neurogenesis, tubulin biology, ... [more]

Status: Preliminary Report

Quantitative Score

  • 0.985542202 [compPASS Score]

Throughput

  • High Throughput

Additional Notes

  • APMS data generated in HEK293T by overexpression of Strep-tagged patient derived ASD variant. Intensity of the interaction between the WT protein and the hit protein was compared to the intensity of the interaction between the patient derived variant and hit protein. Significant interaction intensity changes were defined as those with a log2(fold-change) >= 1 and p <= 0.05. Strengthened interactions were denoted as 'Up', weakened interactions were denoted as 'Down' and unchanged interactions were denoted as 'Not significant' based on these scores. ASD variant: Bait protein name: G64D - Interaction Strength: not significant, Log2fc: 1.5089330348357, p-value: 0.0799087454111538
  • APMS data generated in HEK293T by overexpression of Strep-tagged patient derived ASD variant. Intensity of the interaction between the WT protein and the hit protein was compared to the intensity of the interaction between the patient derived variant and hit protein. Significant interaction intensity changes were defined as those with a log2(fold-change) >= 1 and p <= 0.05. Strengthened interactions were denoted as 'Up', weakened interactions were denoted as 'Down' and unchanged interactions were denoted as 'Not significant' based on these scores. ASD variant: Bait protein name: R10W - Interaction Strength: not significant, Log2fc: 0.809731573735643, p-value: 0.292457220506493
  • Affinity purification-MS was used to capture high confidence wild-type protein interactors as defined by the thresholds: BFDR <= 0.05 and compPASS scores (rank_WD) >= 0.971. Baits are Strep-tagged proteins exogenously expressed in HEK 293T cells. ComPASS score: 0.985542202

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
AP2M1 AP2S1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High-BioGRID
726625
AP2S1 AP2M1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High-BioGRID
3347877
AP2M1 AP2S1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High1BioGRID
1194196
AP2S1 AP2M1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High1BioGRID
2216535
AP2S1 AP2M1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High1BioGRID
3122731
AP2M1 AP2S1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High1BioGRID
3179607
AP2M1 AP2S1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

Low/High-BioGRID
938436
AP2M1 AP2S1
Co-fractionation
Co-fractionation

Interaction inferred from the presence of two or more protein subunits in a partially purified protein preparation. If co-fractionation is demonstrated between 3 or more proteins, then add them as a complex.

High-BioGRID
3789267
AP2S1 AP2M1
Co-fractionation
Co-fractionation

Interaction inferred from the presence of two or more protein subunits in a partially purified protein preparation. If co-fractionation is demonstrated between 3 or more proteins, then add them as a complex.

High0.5091BioGRID
1258650
AP2M1 AP2S1
Cross-Linking-MS (XL-MS)
Cross-Linking-MS (XL-MS)

An interaction is detected between two proteins using chemically reactive or photo-activatable cross-linking reagents that covalently link amino acids in close proximity, followed by mass spectrometry analysis to identify the linked peptides (reviewed in PMID 37406423, 37104977). Experiments may be carried with live cells or cell lysates in which all proteins are expressed at endogenous levels (e.g. PMID 34349018, 35235311) or with recombinant proteins (e.g., PMID 28537071).

High-BioGRID
3792924

Curated By

  • BioGRID