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You are negatively free to the extent that other people do not restrict what you can do. Negative freedom is freedom from interference. The 20th-century political philosopher Isaiah Berlin (1909-97) thought that the answer to both these questions was ‘Yes’, and in his essay ‘Two Concepts of Liberty’ (1958) he distinguished two kinds of freedom (or liberty Berlin used the words interchangeably), which he called negative freedom and positive freedom. Might there be different kinds of freedom and, if so, could the different kinds conflict with each other? Could the promotion of one kind of freedom limit another kind? Could people even be coerced in the name of freedom? But what exactly do we mean by ‘freedom’? The fact that politicians of all parties claim to believe in freedom suggests that people don’t always have the same thing in mind when they talk about it. We all respond positively to it, and under its banner revolutions have been started, wars have been fought, and political campaigns are continually being waged.
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Read more about how to correctly acknowledge RSC content.‘Freedom’ is a powerful word. Permission is not required) please go to the Copyright If you want to reproduce the wholeĪrticle in a third-party commercial publication (excluding your thesis/dissertation for which If you are the author of this article, you do not need to request permission to reproduce figuresĪnd diagrams provided correct acknowledgement is given. Provided correct acknowledgement is given. If you are an author contributing to an RSC publication, you do not need to request permission Please go to the Copyright Clearance Center request page. To request permission to reproduce material from this article in a commercial publication, Provided that the correct acknowledgement is given and it is not used for commercial purposes. This article in other publications, without requesting further permission from the RSC, Hackenberger,Ĭreative Commons Attribution-NonCommercial 3.0 Unported Licence. We believe this integrated approach can be widely adopted to study the structure and interaction of poorly characterized enzyme–substrate complexes, in particular with synthetically challenging or labile substrates.Ĭombining free energy calculations with tailored enzyme activity assays to elucidate substrate binding of a phospho-lysine phosphataseĪ. Comparison of simulations with experiments clearly suggested a distinct binding motif of pLys peptides as well as a very narrow promiscuity of LHPP. Molecular dynamics based free energy calculations, which are unique in their accuracy and solid theoretical basis, were further applied to predict relative binding affinity of different substrates. Molecular docking was employed to explore possible binding poses of the substrates in complex with the enzyme. Taking the instability of the phosphoramidate bond into account, we conducted a carefully adjusted enzymatic assay with various pLys pentapeptides to confirm enzymatic phosphatase activity with LHPP. LHPP has been previously described to hydrolyze the phosphoramidate bonds in different small molecule substrates, including phosphorylated lysine (pLys). In this article, we elucidate the enzymatic activity of Phospholysine Phosphohistidine Inorganic Pyrophosphate Phosphatase (LHPP) towards different O- and N-phosphorylated peptides by a combined experimental and computational approach. Such investigations can be particularly challenging when the modification itself is intrinsically labile. Studying enzymes that are involved in the regulation of dynamic post-translational modifications (PTMs) is of key importance in proteomics research.