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Results: 4

1.
Fig. 1.

Fig. 1. From: Streamlined analysis schema for high-throughput identification of endogenous protein complexes.

IP/MS optimization for deep interactome coverage. (A) Immunoprecipitation procedure for purification of extended endogenous complexes. (B) Proteins in IP/MS result can be separated into the specific and nonspecific categories. Specific proteins constitute antibody affinities, including targeted (intended) and nontargeted (secondary, cross-reacting) complexes.

Anna Malovannaya, et al. Proc Natl Acad Sci U S A. 2010 February 9;107(6):2431-2436.
2.
Fig. 2.

Fig. 2. From: Streamlined analysis schema for high-throughput identification of endogenous protein complexes.

Extended Integrator interactome. (A) Reciprocal IPs against Integrator subunits retrieve previously known core module and interacting polymerase subunits. (B) Multiple new interactors are discovered consistently with the Integrator: a phosphatase module, OBFC2A/B, four uncharacterized predicted proteins, and a unique Z3 complex consisting of ZMYND8, ZNF687, and ZNF592. (C) Reproducible antibody-specific identifications contain potential antibody cross-reactivity.

Anna Malovannaya, et al. Proc Natl Acad Sci U S A. 2010 February 9;107(6):2431-2436.
3.
Fig. 4.

Fig. 4. From: Streamlined analysis schema for high-throughput identification of endogenous protein complexes.

De novo IP/MS deconvolution of human HDAC1/2 corepressor complex network. (A) HDAC1-containing CHD4, SIN3A, and RCOR1 complexes were defined by comparison of reciprocal 3Ns. Heterogeneity of HDAC1/2 complexes is revealed as these modules break apart from each other in 3N analysis. Proteins that were directly targeted as antigens are shaded in blue; unique core complex associations are highlighted in orange. (B) Subunit assignment for the HDAC1/2 network intercomplex interactor PBRM1/BRD7 complex.

Anna Malovannaya, et al. Proc Natl Acad Sci U S A. 2010 February 9;107(6):2431-2436.
4.
Fig. 3.

Fig. 3. From: Streamlined analysis schema for high-throughput identification of endogenous protein complexes.

Core complex subunits of Mediator are defined by 3N analysis. (A) Top IPs where MED12 is present at highest levels (> 5 peptides) were clustered with 3N constraints (see text). BL#, antibody IDs; ∗ identifies primary antibodies where Mediator is a secondary interacting or cross-reacting complex. (B) 3N analysis was performed for all Mediator subunits with sufficient number of identification in our dataset. Protein neighbors that are copresent in multiple reciprocal 3Ns (•) define potential core complex clusters for Mediator. Mediator-interacting polymerase is effectively stratified from the Mediator core by this analysis.

Anna Malovannaya, et al. Proc Natl Acad Sci U S A. 2010 February 9;107(6):2431-2436.

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