1,721,008 research outputs found
PDZ domains: folding and binding.
No full textThe PDZ domain is one of the most common protein-protein interaction domains in humans, and it is found in all kingdoms of life. We will review recent progress in the understanding of biophysical aspects of PDZ domains with emphasis on the folding and binding reactions. Finally, we discuss an intriguing correlation between stability and binding of peptide for PDZ2 from PTP-BL.</p
Supertertiary protein structure affects an allosteric network
Full text linkThe notion that protein function is allosterically regulated by structural or dynamic changes in proteins has been extensively investigated in several protein domains in isolation. In particular, PDZ domains have represented a paradigm for these studies, despite providing conflicting results. Furthermore, it is still unknown how the association between protein domains in supramodules, consitituting so-called supertertiary structures, affects allosteric networks. Here, we experimentally mapped the allosteric network in a PDZ:ligand complex, both in isolation and in the context of a supramodular structure, and show that allosteric networks in a PDZ domain are highly dependent on the supertertiary structure in which they are present. This striking sensitivity of allosteric networks to the presence of adjacent protein domains is likely a common property of supertertiary structures in proteins. Our findings have general implications for prediction of allosteric networks from primary and tertiary structures and for quantitative descriptions of allostery
Seeking allosteric networks in PDZ domains
Full text linkEver since Ranganathan and coworkers subjected the covariation of amino acid residues in the postsynaptic density-95/Discs large/Zonula occludens 1 (PDZ) domain family to a statistical correl- ation analysis, PDZ domains have represented a paradigmatic family to explore single domain protein allostery. Nevertheless, several theoretical and experimental studies in the past two dec- ades have contributed contradicting results with regard to structural localization of the allosteric networks, or even questioned their actual existence in PDZ domains. In this review, we first describe theoretical and experimental approaches that were used to probe the energetic network (s) in PDZ domains. We then compare the proposed networks for two well-studied PDZ domains namely the third PDZ domain from PSD-95 and the second PDZ domain from PTP-BL. Our analysis highlights the contradiction between the different methods and calls for additional work to better understand these allosteric phenomena
Coupled binding and folding of intrinsically disordered proteins: what can we learn from kinetics?
No full textProtein or protein regions that are not forming well-defined
structures in their free states under native-like conditions are
called intrinsically disordered proteins. Such proteins are very
common in protein–protein interactions, where their disorder
apparently gives several advantages including optimal binding
properties. To fully appreciate why protein disorder is
advantageous for protein–protein interactions we need to
understand the mechanism(s) of interaction. However,
elucidating mechanisms in protein–protein interactions is
usually very challenging. Here we discuss how kinetics in
combination with protein engineering and structural
information can be used to depict details of protein–protein
interactions involving intrinsically disordered proteins
The binding mechanisms of intrinsically disordered proteins
Full text linkIntrinsically disordered proteins (IDPs) and intrinsically disordered regions (IDRs) of proteins are very common and instrumental for cellular signaling. Recently, a number of studies have investigated the kinetic binding mechanisms of IDPs and IDRs. These results allow us to draw conclusions about the energy landscape for the coupled binding and folding of disordered proteins. The association rate constants of IDPs cover a wide range (10(5)-10(9) M-1 s(-1)) and are largely governed by long-range charge-charge interactions, similarly to interactions between well-folded proteins. Off-rate constants also differ significantly among IDPs (with half-lives of up to several minutes) but are usually around 0.1-1000 s(-1), allowing for rapid dissociation of complexes. Likewise, affinities span from pM to mu M suggesting that the low-affinity high-specificity concept for IDPs is not straightforward. Overall, it appears that binding precedes global folding although secondary structure elements such as helices may form before the protein-protein interaction. Short IDPs bind in apparent two-state reactions whereas larger IDPs often display complex multi-step binding reactions. While the two extreme cases of two-step binding (conformational selection and induced fit) or their combination into a square mechanism is an attractive model in theory, it is too simplistic in practice. Experiment and simulation suggest a more complex energy landscape in which IDPs bind targets through a combination of conformational selection before binding (e. g., secondary structure formation) and induced fit after binding (global folding and formation of short-range intermolecular interactions)
Understanding the role of phosphorylation in the binding mechanism of a PDZ domain
No full textThe PDZ domain is one of the most common protein–protein interaction domains in mammalian species. While several studies have demonstrated the importance of phosphorylation in interactions involving PDZ domains, there is a paucity of detailed mechanistic data addressing how the PDZ interaction is affected by phosphorylation. Here, we address this question by equilibrium and
kinetic binding experiments using PDZ2 from protein tyrosine phosphatase L1 and its interaction with a peptide from the natural ligand RIL. The results show that phosphorylation of a serine residue in the RIL peptide has dual and opposing effects: it increases both the association and dissociation rate constants, which leads to an overall weakening of binding. Furthermore, we performed
binding experiments with a RIL peptide in which the serine was replaced by a glutamate, a commonly used method to mimic phosphorylation in proteins. Strikingly, both the affinity and the ionic strength dependence of the affinity differed markedly for the phosphoserine and glutamate peptides. These results show that, in this particular case, glutamate is a poor mimic of serine
phosphorylation
Simulation and Experiment Conspire to reveal Cryptic Intermediates and the Slide from Framework to Nucleation-Condensation Mechanism in the Folding of the Homeodomain Superfamily.
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