A Hot Spot of Binding Energy in a Hormone-Receptor Interface

Clackson, Tim; Wells, James A.

doi:10.1126/science.7529940

Cited by 1,930 publications

(1,640 citation statements)

References 37 publications

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“…To produce this recognition layer by following a protein mimetic approach, we selected organosilanes that mimic the lateral chains of amino acids known to be of importance in protein-protein interactions (Fig. 1c) 33,34 . We hypothesized that these building blocks will self-assemble at the surface of the virions before their covalent incorporation within the recognition layer.…”

Section: Resultsmentioning

confidence: 99%

A synthetic nanomaterial for virus recognition produced by surface imprinting

et al. 2013

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Major stumbling blocks in the production of fully synthetic materials designed to feature virus recognition properties are that the target is large and its self-assembled architecture is fragile. Here we describe a synthetic strategy to produce organic/inorganic nanoparticulate hybrids that recognize non-enveloped icosahedral viruses in water at concentrations down to the picomolar range. We demonstrate that these systems bind a virus that, in turn, acts as a template during the nanomaterial synthesis. These virus imprinted particles then display remarkable selectivity and affinity. The reported method, which is based on surface imprinting using silica nanoparticles that act as a carrier material and organosilanes serving as biomimetic building blocks, goes beyond simple shape imprinting. We demonstrate the formation of a chemical imprint, comparable to the formation of biosilica, due to the template effect of the virion surface on the synthesis of the recognition material.

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Section: Resultsmentioning

confidence: 99%

A synthetic nanomaterial for virus recognition produced by surface imprinting

et al. 2013

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“…As mentioned above, for Site-1, the major hGHbp binding determinants are two loops, each containing an essential tryptophan residue. On binding, the tryptophan side chains become deeply buried in the interface and act as essential anchors for productive binding (7). Importantly, it has been shown by X-ray structural analysis that the packing of the W104 and W169 side chains in complexes with wt-hGH and hGHv are virtually identical (5).…”

Section: (I) What Happened To the Binding Hot-spot?mentioning

confidence: 99%

“…Mutations in hGHbp that remove either of the Trp residues are extremely deleterious to binding of wt-hGH, resulting in a 10 000-fold reduction in the binding constant (7). However, for hGHv, the same mutations produce a drastically reduced effect; the W104A and W169A mutants bind with K d values of 2.3 and 9.2 nM, respectively (∼400-1000-fold reduction in affinity).…”

Section: (I) What Happened To the Binding Hot-spot?mentioning

confidence: 99%

The Role of Protein Dynamics in Increasing Binding Affinity for an Engineered Protein−Protein Interaction Established by H/D Exchange Mass Spectrometry

et al. 2006

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It is generally accepted that protein and solvation dynamics play fundamental roles in the mechanisms of protein-protein binding; however, assessing their contribution meaningfully has not been straightforward. Here, hydrogen/deuterium exchange mass spectrometry (H/D-Ex) was employed to assess the role of dynamics for a high-affinity human growth hormone variant (hGHv) and the wild-type growth hormone (wt-hGH) each binding to the extracellular domain of their receptor (hGHbp). Comparative analysis of the transient fluctuations in the bound and unbound states revealed that helix-1 of hGHv undergoes significant transient unfolding in its unbound state, a characteristic that was not found in wthGH or apparent in the temperature factor data from the X-ray analysis of the unbound hGHv structure. In addition, upon hormone binding, an overall increase in stability was observed for the -sheet structure of hGHbp which included sites distant from the binding interface. On the basis of the stability, binding kinetics, and thermodynamic data presented, the increase in the binding free energy of hGHv is primarily generated by factors that appear to increase the energy of the unbound state relative to the free energy of the bound complex. This implies that an alternate route to engineer new interactions aiming to increase protein-protein association energies may be achieved by introducing certain mutations that destabilize one of the interacting molecules without destabilizing the resulting bound complex. Importantly, although the hGHv molecule is less stable than its wt-hGH counterpart, its resulting active ternary complex with two copies of hGHbp has comparable stability to the wt complex.A principal goal in structural biophysics is to develop general rules for structure-function relationships that govern protein-protein interactions, ultimately to quantify the contribution of individual local interactions to the overall binding energy. In this regard, there is a growing appreciation that essentially all interactions are context-dependent, and factors such as solvation and dynamics play critical, if not dominant, roles in forming most surface-to-surface contacts. Because solvation and dynamics are inherently difficult phenomena to quantify experimentally, their influence many times is invoked in a very general way, especially when structure-based rationales fail to explain biophysical data.The interaction between a high-affinity variant of human growth hormone (hGH) and the extracellular domain of its cognate receptor (hGHbp) 1 represents one of the cases where the binding properties are difficult to rationalize based on general structure-function principals. This variant (hGHv) was produced by phage display mutagenesis and binds through Site-1 to hGHbp with a K d < 10 pM, which constitutes a ∼400-fold (∼3 kcal/mol) increased affinity over the wild-type hormone (wt-hGH), K d ∼1 nM (1-3). It contains 15 mutations in its Site-1 binding interface and retains its full biological activity (4).An unanticipated finding w...

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“…More detailed analyses of the contribution of individual residues to the binding sites of both GH and its receptor have been undertaken (Cunningham et al, 1991; Cunningham & Wells, 1993;Clackson & Wells, 1995). In these studies, alanine scanning mutagenesis was performed on both GH and the GHR and their binding affinities, relative to wild-type, were calculated.…”

Section: The Complex Of Growth Hormone and Its Receptormentioning

confidence: 99%

Interleukin‐6: Structure‐function relationships

et al. 1997

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Interleukin-6 (IL-6) is a multifunctional cytokine that plays a central role in host defense due to its wide range of immune and hematopoietic activities and its potent ability to induce the acute phase response. Overexpression of IL-6 has been implicated in the pathology of a number of diseases including multiple myeloma, rheumatoid arthritis, Castleman's disease, psoriasis, and post-menopausal osteoporosis. Hence, selective antagonists of IL-6 action may offer therapeutic benefits. IL-6 is a member of the family of cytokines that includes interleukin-1 1, leukemia inhibitory factor, oncostatin M, cardiotrophin-1, and ciliary neurotrophic factor. Like the other members of this family, IL-6 induces growth or differentiation via a receptor-system that involves a specific receptor and the use of a shared signaling subunit, gp130. Identification of the regions of IL-6 that are involved in the interactions with the IL-6 receptor and gp130 is an important first step in the rational manipulation of the effects of this cytokine for therapeutic benefit. In this review, we focus on the sites on IL-6 which interact with its low-affhity specific receptor, the IL-6 receptor, and the high-affinity converter gp130. A tentative model for the IL-6 hexameric receptor ligand complex is presented and discussed with respect to the mechanism of action of the other members of the IL-6 family of cytokines.

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A Hot Spot of Binding Energy in a Hormone-Receptor Interface

Cited by 1,930 publications

References 37 publications

A synthetic nanomaterial for virus recognition produced by surface imprinting

A synthetic nanomaterial for virus recognition produced by surface imprinting

The Role of Protein Dynamics in Increasing Binding Affinity for an Engineered Protein−Protein Interaction Established by H/D Exchange Mass Spectrometry

Interleukin‐6: Structure‐function relationships

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