Thermodynamic study of the BRCT domain of BARD1 and its interaction with the -pSER-X-X-Phe- motif-containing BRIP1 peptide.


The BRCA1-associated RING domain protein 1 (BARD1) is the heterodimeric partner of BRCA1. The BRCA1/BARD1 complex demonstrates ubiquitin ligase activity and has been implicated in genomic stability and tumor suppression. Both proteins possess a structurally conserved C-terminal domain (BRCT). While BRCA1-BRCT has been shown to mediate BRCA1 interactions with phosphoproteins such as BRIP1 by recognizing the pSer-X-X-Phe motif, attempts to demonstrate analogous interactions of its dimeric counterpart BARD1-BRCT, have so far been unsuccessful. In this study, chemical-denaturation experiments of BARD1-BRCT domain suggest that its low thermodynamic stability (DeltaG=2.5 kcal/mol) at room temperature, may affect some of its biochemical properties, such as its interaction with phosphopeptides. The stability of BARD1-BRCT domain at 10 degrees C, increases to 7.5 kcal/mol and isothermal titration calorimetry (ITC) experiments at this lower temperature showed binding to the BRIP1 phosphopeptide via an enthalpy-driven interaction, which appears to be specific to the pSer-X-X-Phe peptide-binding motif. Substitution of either pSer at position 0 with Ser (non-phosphorylated peptide) or Phe with Val at position +3, leads to no-binding ITC results. While these findings are indicative that BRIP1 is a potential BARD1 binding partner, it becomes evident that in vitro binding assays involving the entire BARD1 protein and in vivo experiments are also needed to establish its binding partners and its potential role in tumor suppression pathways. Study holds ProTherm entries: 25716, 25717, 25718, 25719 Extra Details: BRAC1: Breast Cancer gene 1 protein; conserved C-terminal domain; 6xHis-tag Chemical unfolding; Thermodynamic stability; ITC binding experiments

Submission Details


Submitter: Connie Wang

Submission Date: April 24, 2018, 8:56 p.m.

Version: 1

Publication Details
Thanassoulas A;Nomikos M;Theodoridou M;Yannoukakos D;Mastellos D;Nounesis G,Biochim. Biophys. Acta (2010) Thermodynamic study of the BRCT domain of BARD1 and its interaction with the -pSER-X-X-Phe- motif-containing BRIP1 peptide. PMID:20451671
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1JM7 2001-07-17T00:00:00+0000 0 Solution structure of the BRCA1/BARD1 RING-domain heterodimer
2NTE 2006-11-07T00:00:00+0000 1.9 Crystal Structure of the BARD1 BRCT Domains
2R1Z 2007-08-23T00:00:00+0000 2.1 Crystal Structure of the BARD1 BRCT Repeat
3C5R 2008-02-01T00:00:00+0000 2.0 Crystal Structure of the BARD1 Ankyrin Repeat Domain and Its Functional Consequences
3FA2 2008-11-14T00:00:00+0000 2.2 Crystal Structure of the BRCA1 Associated Ring Domain (BARD1) Tandem BRCT Domains
6M14 2020-02-24T00:00:00+0000 1.88 Crystal Structure of the BARD1 BRCT Mutant
7E8I 2021-03-01T00:00:00+0000 3.1 Structural insight into BRCA1-BARD1 complex recruitment to damaged chromatin
7JZV 2020-09-02T00:00:00+0000 3.9 Cryo-EM structure of the BRCA1-UbcH5c/BARD1 E3-E2 module bound to a nucleosome
7LYB 2021-03-06T00:00:00+0000 3.28 Cryo-EM structure of the human nucleosome core particle in complex with BRCA1-BARD1-UbcH5c
7LYC 2021-03-06T00:00:00+0000 2.94 Cryo-EM structure of the human nucleosome core particle ubiquitylated at histone H2A Lys13 and Lys15 in complex with BARD1 (residues 415-777)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 BRCA1-associated RING domain protein 1 Q99728 BARD1_HUMAN