Structure of the CCR5 Chemokine Receptor–HIV Entry Inhibitor Maraviroc Complex

Tan, Qiuxiang; Zhu, Yun; Li, Jian; Chen, Zhuxi; Han, Gye Won; Kufareva, Irina; Li, Tingting; Ma, Limin; Fenalti, Gustavo; Li, Jing; Zhang, Wenru; Xie, Xin; Yang, Huaiyu; Jiang, Hualiang; Cherezov, Vadim; Liu, Hong; Stevens, Raymond C.; Zhao, Qiang; Wu, Beili

doi:10.1126/science.1241475

Cited by 631 publications

(760 citation statements)

References 35 publications

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“…Finding that two similar but distinct receptors bind the same chemokine, and that CXCR1 almost exclusively binds to IL-8 whereas CXCR2 also binds to several other chemokines, provided an early indication of the complexity of the chemokine defense system. At present, the structures of six chemokine receptors have been reported: the crystal structures of CXCR4 (24,25), CCR2 (26), CCR5 (27), CCR9 (28), and viral US28 (29), and our NMR structure of CXCR1 in lipid bilayers (23), which is shown in Fig. 1 B. Although none of the structures represent complexes of a receptor with one of its native chemokine agonists, the structures of receptors bound to viral chemokines provide information that is complementary to the many mutagenesis and binding studies of their interactions.…”

Section: Introductionmentioning

confidence: 99%

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Park

Berkamp

Radoicic

et al. 2017

Biophysical Journal

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The human chemokine interleukin-8 (IL-8; CXCL8) is a key mediator of innate immune and inflammatory responses. This small, soluble protein triggers a host of biological effects upon binding and activating CXCR1, a G proteincoupled receptor, located in the cell membrane of neutrophils. Here, we describe 1 H-detected magic angle spinning solid-state NMR studies of monomeric IL-8 (1-66) bound to full-length and truncated constructs of CXCR1 in phospholipid bilayers under physiological conditions. Cross-polarization experiments demonstrate that most backbone amide sites of IL-8 (1-66) are immobilized and that their chemical shifts are perturbed upon binding to CXCR1, demonstrating that the dynamics and environments of chemokine residues are affected by interactions with the chemokine receptor. Comparisons of spectra of IL-8 (1-66) bound to full-length CXCR1 (1-350) and to N-terminal truncated construct NT-CXCR1 (39-350) identify specific chemokine residues involved in interactions with binding sites associated with N-terminal residues (binding site-I) and extracellular loop and helical residues (binding site-II) of the receptor. Intermolecular paramagnetic relaxation enhancement broadening of IL-8 (1-66) signals results from interactions of the chemokine with CXCR1 (1-350) containing Mn 2þ chelated to an unnatural amino acid assists in the characterization of the receptor-bound form of the chemokine.

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Section: Introductionmentioning

confidence: 99%

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Park

Berkamp

Radoicic

et al. 2017

Biophysical Journal

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“…This unique position suggests their role as signal initiators responsive to chemokine binding. One of these CXCR4 residues, W94 2.60 , is highly conserved among chemokine receptors, has been implicated in binding the small molecule antagonist IT1t and vMIP-II (8,9), and the equivalent residue in the chemokine receptor CCR5 (W86 2.60 ) has also been shown to play a role in ligand binding (24). Additional signal initiator residues identified in our screen include Y45 1.39 , Y116 3.32 , and E288 7.39 , all previously reported as binding and/ or signaling determinants in CXCR4 (13,19,22,25).…”

Section: Significancementioning

confidence: 99%

Signal transmission through the CXC chemokine receptor 4 (CXCR4) transmembrane helices

Wescott

Kufareva

Paes

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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183

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The atomic-level mechanisms by which G protein-coupled receptors (GPCRs) transmit extracellular ligand binding events through their transmembrane helices to activate intracellular G proteins remain unclear. Using a comprehensive library of mutations covering all 352 residues of the GPCR CXC chemokine receptor 4 (CXCR4), we identified 41 amino acids that are required for signaling induced by the chemokine ligand CXCL12 (stromal cell-derived factor 1). CXCR4 variants with each of these mutations do not signal properly but remain folded, based on receptor surface trafficking, reactivity to conformationally sensitive monoclonal antibodies, and ligand binding. When visualized on the structure of CXCR4, the majority of these residues form a continuous intramolecular signaling chain through the transmembrane helices; this chain connects chemokine binding residues on the extracellular side of CXCR4 to G proteincoupling residues on its intracellular side. Integrated into a cohesive model of signal transmission, these CXCR4 residues cluster into five functional groups that mediate (i) chemokine engagement, (ii) signal initiation, (iii) signal propagation, (iv) microswitch activation, and (v) G protein coupling. Propagation of the signal passes through a "hydrophobic bridge" on helix VI that coordinates with nearly every known GPCR signaling motif. Our results agree with known conserved mechanisms of GPCR activation and significantly expand on understanding the structural principles of CXCR4 signaling.T he CXC chemokine receptor 4 (CXCR4) belongs to the G protein-coupled receptor (GPCR) superfamily of proteins, the largest class of integral membrane proteins encoded in the human genome, comprising greater than 30% of current drug targets. Deregulation of CXCR4 expression in multiple human cancers, its role in hematopoietic stem cell migration, and the utilization of CXCR4 by HIV-1 for T-cell entry, make this receptor an increasingly important therapeutic target (1). One FDA-approved drug against CXCR4 is currently on the market (Mozobil, for hematopoietic stem cell mobilization), and multiple additional drugs against this target are in development for oncology and other indications (2).The crystal structures of class A GPCR superfamily members in their active and inactive conformations (reviewed in refs. 3 and 4) provide unprecedented insight into the structural basis of ligand binding, G protein coupling, and activation of GPCRs via rearrangements of transmembrane (TM) helices. GPCR helices V and VI in particular, and in some cases III and VII, are known to undergo significant conformational changes upon activation (5-7). However, static images alone have not been able to explain the residue-level mechanisms underlying the dynamic helical shifts that mediate GPCR signal transduction. Additionally, only inactive state structures have been solved for CXCR4 and most other GPCRs (8,9). Over the last two decades, extensive mutagenesis studies of GPCRs in general [collectively describing >8,000 mutations (gpcrdb.org)] and of CX...

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“…In addition, researchers routinely add fusion partners such as T4 lysozyme or apocytochrome b 562 RIL to either the N terminus or the second or third intracellular loops (11). Other fusion partners used include the catalytic domain of Pyrococcus abyssi glycogen synthase in the orexin 2 receptor (12) and rubredoxin in CCR5 (13). These fusion partners are selected because they are stable domains that crystallize readily, but more importantly, their N and C termini are in close proximity (less than 15 Å), thus allowing them to be inserted in the receptor loops without gross alteration of the receptor structure.…”

Section: Overview Of Technology Developments Enabling Gpcr Structuresmentioning

confidence: 99%

From G Protein-coupled Receptor Structure Resolution to Rational Drug Design

Jazayeri

Dias

Marshall

2015

Journal of Biological Chemistry

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A number of recent technical solutions have led to significant advances in G protein-coupled receptor (GPCR) structural biology. Apart from a detailed mechanistic view of receptor activation, the new structures have revealed novel ligand binding sites. Together, these insights provide avenues for rational drug design to modulate the activities of these important drug targets. The application of structural data to GPCR drug discovery ushers in an exciting era with the potential to improve existing drugs and discover new ones. In this review, we focus on technical solutions that have accelerated GPCR crystallography as well as some of the salient findings from structures that are relevant to drug discovery. Finally, we outline some of the approaches used in GPCR structure based drug design.The process of drug discovery from bench to market is a high risk long term investment that has been estimated to cost about $1.8 billion per drug (1). Such high costs coupled with the pressure of patent expiry dates and increased regulatory constraints have propelled the pharmaceutical industry to increase efficiency of research and development to reduce the attrition rate. A key area of focus has been improvements in the quality of compounds that are discovered in the early stages of the drug discovery pipeline. The main driver for this effort has been the general observation that the hits identified from cell-based high throughput screening (HTS) 2 strategies are usually large lipophilic molecules that are difficult to optimize and carry a number of liabilities that significantly increase their failure rate (2). One of the main advances to tackle these issues has been the utilization of fragment-based drug discovery (FBDD) approaches that rely on screening small chemical fragments (100 -250 Da). Because of their small size, a significantly larger portion of chemical space can be explored with fewer compounds when compared with HTS. In addition, initial hits from a fragment screen bind more efficiently to their target and represent excellent starting points for medicinal chemists to grow and optimize these into lead and candidate molecules (3). The initial fragment hits exhibit low affinity, so they need to be screened at high concentrations that make them incompatible with biological assays. Instead, biophysical assays are used in FBDD cascades, and in addition, these approaches are combined with structural information derived primarily from x-ray crystallography. The application of structure-based drug design (SBDD) allows medicinal chemists to rationally convert fragment hits into larger compounds with higher affinity while maintaining the efficiency of binding and drug-like properties. Historically, SBDD methods gained traction with soluble proteins such as enzymes as routine generation of structural data is facilitated by their high stability in purified form (4 -6). This is in sharp contrast to membrane proteins such as G proteincoupled receptors (GPCRs) that have been refractory to SBDD due to the challenges associate...

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Structure of the CCR5 Chemokine Receptor–HIV Entry Inhibitor Maraviroc Complex

Cited by 631 publications

References 35 publications

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Signal transmission through the CXC chemokine receptor 4 (CXCR4) transmembrane helices

From G Protein-coupled Receptor Structure Resolution to Rational Drug Design

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