Two Bacterial Small Heat Shock Proteins, IbpA and IbpB, Form a Functional Heterodimer.
Small heat shock proteins (sHsps) are a conserved class of ATP-independent chaperones which in stress conditions bind to unfolded protein substrates and prevent their irreversible aggregation. Substrates trapped in sHsps-containing aggregates are efficiently refolded into native structures by ATP-dependent Hsp70 and Hsp100 chaperones. Most γ-proteobacteria possess a single sHsp (IbpA), while in a subset of Enterobacterales, as a consequence of ibpA gene duplication event, a two-protein sHsp (IbpA and IbpB) system has evolved. IbpA and IbpB are functionally divergent. Purified IbpA, but not IbpB, stably interacts with aggregated substrates, yet both sHsps are required to be present at the substrate denaturation step for subsequent efficient Hsp70-Hsp100-dependent substrate refolding. IbpA and IbpB interact with each other, influence each other's expression levels and degradation rates. However, the crucial information on how these two sHsps interact and what is the basic building block required for proper sHsps functioning was missing. Here, based on NMR, mass spectrometry and crosslinking studies, we show that IbpA-IbpB heterodimer is a dominating functional unit of the two sHsp system in Enterobacterales. The principle of heterodimer formation is similar to one described for homodimers of single bacterial sHsps. β-hairpins formed by strands β5 and β7 of IbpA or IbpB crystallin domains associate with the other one's β-sandwich in the heterodimer structure. Relying on crosslinking and molecular dynamics studies, we also propose the orientation of two IbpA-IbpB heterodimers in a higher order tetrameric structure.
['Piróg|Artur|A|', 'Cantini|Francesca|F|', 'Nierzwicki|Łukasz|Ł|', 'Obuchowski|Igor|I|', 'Tomiczek|Bartłomiej|B|', 'Czub|Jacek|J|', 'Liberek|Krzysztof|K|']
D001426:Bacterial Proteins D001665:Binding Sites D004755:Enterobacteriaceae D050888:Heat-Shock Proteins, Small D009682:Magnetic Resonance Spectroscopy D013058:Mass Spectrometry D008958:Models, Molecular D056004:Molecular Dynamics Simulation D011485:Protein Binding D000072757:Protein Conformation, beta-Strand D055503:Protein Multimerization
bacterial small heat shock proteins chaperones heterodimer of sHsps modification of protein aggregation protein refolding