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

Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.

Huttlin EL, Bruckner RJ, Navarrete-Perea J, Cannon JR, Baltier K, Gebreab F, Gygi MP, Thornock A, Zarraga G, Tam S, Szpyt J, Gassaway BM, Panov A, Parzen H, Fu S, Golbazi A, Maenpaa E, Stricker K, Guha Thakurta S, Zhang T, Rad R, Pan J, Nusinow DP, Paulo JA, Schweppe DK, Vaites LP, Harper JW, Gygi SP

Thousands of interactions assemble proteins into modules that impart spatial and functional organization to the cellular proteome. Through affinity-purification mass spectrometry, we have created two proteome-scale, cell-line-specific interaction networks. The first, BioPlex 3.0, results from affinity purification of 10,128 human proteins-half the proteome-in 293T cells and includes 118,162 interactions among 14,586 proteins. The second results from 5,522 immunoprecipitations in HCT116 ... [more]

Cell May. 27, 2021; 184(11);3022-3040.e28 [Pubmed: 33961781]

Quantitative Score

  • 0.999994341 [compPASS Score]

Throughput

  • High Throughput

Additional Notes

  • BioPlex 3.0 HEK 293T cells CompPASS score = 0.999994341, threshold = 0.75. Quantitative scores are calculated by CompPASS-Plus (Huttlin et al. Cell 2015, PMID: 26186194). The 0.75 threshold represents the top 2% of scores in HEK293T.
  • BioPlex HCT HCT116 cells CompPASS score = 0.999999896, threshold = 0.75. Quantitative scores are calculated by CompPASS-Plus (Huttlin et al. Cell 2015, PMID: 26186194). The 0.75 threshold represents the top 2% of scores in HCT116.
  • Only scores from within the same cell line in BioPlex HCT (PMID: 33961781) should be compared directly. For comparison of HEK293T and HCT116 interaction networks with relaxed threshold = 0.1, see BioPlex Interactome (https://bioplex.hms.harvard.edu/index.php).
  • This data may be re-scored from BioPlex 1.0 (PMID: 26186194) and BioPlex 2.0 (PMID: 28514442). Only scores from within the same cell line in BioPlex 3.0 (PMID: 33961781) should be compared directly. For comparison of HEK293T and HCT116 interaction networks with relaxed threshold = 0.1, see BioPlex Interactome (https://bioplex.hms.harvard.edu/index.php).

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
GATAD2A MBD2
Affinity Capture-Luminescence
Affinity Capture-Luminescence

An interaction is inferred when a bait protein, tagged with luciferase, is enzymatically detected in immunoprecipitates of the prey protein as light emission. The prey protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag.

Low-BioGRID
2751956
GATAD2A MBD2
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
2222883
GATAD2A MBD2
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.

High48.7146BioGRID
2942727
MBD2 GATAD2A
Affinity Capture-Western
Affinity Capture-Western

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 identified by Western blot with a specific polyclonal antibody or second epitope tag. This category is also used if an interacting protein is visualized directly by dye stain or radioactivity. Note that this differs from any co-purification experiment involving affinity capture in that the co-purification experiment involves at least one extra purification step to get rid of potential contaminating proteins.

Low-BioGRID
-
GATAD2A MBD2
Affinity Capture-Western
Affinity Capture-Western

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 identified by Western blot with a specific polyclonal antibody or second epitope tag. This category is also used if an interacting protein is visualized directly by dye stain or radioactivity. Note that this differs from any co-purification experiment involving affinity capture in that the co-purification experiment involves at least one extra purification step to get rid of potential contaminating proteins.

Low-BioGRID
-
GATAD2A MBD2
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
3452561
GATAD2A MBD2
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.

High1BioGRID
1264356
MBD2 GATAD2A
Reconstituted Complex
Reconstituted Complex

An interaction is detected between purified proteins in vitro.

Low-BioGRID
-
MBD2 GATAD2A
Reconstituted Complex
Reconstituted Complex

An interaction is detected between purified proteins in vitro.

Low-BioGRID
-
MBD2 GATAD2A
Two-hybrid
Two-hybrid

Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.

High-BioGRID
2724131

Curated By

  • BioGRID