Human CC Research Articles

Structural basis of chemokine recognition by the class A3 tick evasin EVA-ACA1001.

Ticks produce chemokine-binding proteins, known as evasins, in their saliva to subvert the host's immune response. Evasins bind to chemokines and thereby inhibit the activation of their cognate chemokine receptors, thus suppressing leukocyte recruitment and inflammation. We recently described subclass A3 evasins, which, like other class A evasins, exclusively target CC chemokines but appear to use a different binding site architecture to control target selectivity among CC chemokines. We now describe the structural basis of chemokine recognition by the class A3 evasin EVA-ACA1001. EVA-ACA1001 binds to almost all human CC chemokines and inhibits receptor activation. Truncation mutants of EVA-ACA1001 showed that, unlike class A1 evasins, both the N- and C-termini of EVA-ACA1001 play minimal roles in chemokine binding. To understand the structural basis of its broad chemokine recognition, we determined the crystal structure of EVA-ACA1001 in complex with the human chemokine CCL16. EVA-ACA1001 forms backbone-backbone interactions with the CC motif of CCL16, a conserved feature of all class A evasin-chemokine complexes. A hydrophobic pocket in EVA-ACA1001, formed by several aromatic side chains and the unique disulfide bond of class A3 evasins, accommodates the residue immediately following the CC motif (the "CC + 1 residue") of CCL16. This interaction is shared with EVA-AAM1001, the only other class A3 evasins characterized to date, suggesting it may represent a common mechanism that accounts for the broad recognition of CC chemokines by class A3 evasins.

Knowledge, attitudes and practices of cervical cancer screening among female students enrolled in higher education institutions in Cabo Verde.

A descriptive cross-sectional study, using a self-administered structured questionnaire, was conducted in six higher education institutions (HEI) in Cabo Verde between November and December 2020. A total of 618 female undergraduate students were recruited using a simple random sampling technique. Descriptive statistical data analysis was used to report the results. The response rate was 96.6% (n = 618). Most of the participants, 90.6% (549), were single, with average age of 21.79 years (SD =±4). Although most of the participants had already heard about CC (94.6%), most students showed a low knowledge about this disease (86.2%). Moreover, only 9.1% reported having been screened for CC. Most undergraduate female students enrolled in HEI in Cabo Verde have poor knowledge and unfavourable attitudes toward CC. The level of knowledge is quite unsatisfactory. Within this context, the implementation of health policies focused on human papillomavirus education, prevention strategies, and CC screening is crucial.

Systematic Assessment of Human CCR7 Signalling Using NanoBRET Biosensors Points towards the Importance of the Cellular Context.

The human CC chemokine receptor 7 (CCR7) is activated by two natural ligands, CC chemokine ligand 19 (CCL19) and 21 (CCL21). The CCL19-CCL21-CCR7 axis has been extensively studied in vitro, but there is still debate over whether CCL21 is an overall weaker agonist or if the axis displays biased signalling. In this study, we performed a systematic analysis at the transducer level using NanoBRET-based methodologies in three commonly used cellular backgrounds to evaluate pathway and ligand preferences, as well as ligand bias and the influence of the cellular system thereon. We found that both CCL19 and CCL21 activated all cognate G proteins and some non-cognate couplings in a cell-type-dependent manner. Both ligands recruited β-arrestin1 and 2, but the potency was strongly dependent on the cellular system. Overall, CCL19 and CCL21 showed largely conserved pathway preferences, but small differences were detected. However, these differences only consolidated in a weak ligand bias. Together, these data suggest that CCL19 and CCL21 share mostly overlapping, weakly biased, transducer profiles, which can be influenced by the cellular context.

Climate change and geo-environmental factors influencing desertification: a critical review.

The problem of desertification (DSF) is one of the most severe environmental disasters which influence the overall condition of the environment. In Rio de Janeiro Earth Summit on Environment and Development (1922), DSF is defined as arid, semi-arid, and dry sub-humid induced LD and that is adopted at the UNEP's Nairobi ad hoc meeting in 1977. It has been seen that there is no variability in the trend of long-term rainfall, but the change has been found in the variability of temperature (avg. temp. 0-5°C). There is no proof that the air pollution brought on by CO2 and other warming gases is the cause of this rise, which seems to be partially caused by urbanization. The two types of driving factors in DSF-CC (climate change) along with anthropogenic influences-must be compared in order to work and take action to stop DSF from spreading. The proportional contributions of human activity and CC to DSF have been extensively evaluated in this work from "qualitative, semi-quantitative, and quantitative" perspectives. In this study, we have tried to connect the drives of desertification to desertification-induced migration due to loss of biodiversity and agriculture failure. The authors discovered that several of the issues from the earlier studies persisted. The policy-makers should follow the proper SLM (soil and land management) through using the land. The afforestation with social forestry and consciousness among the people can reduce the spreading of the desertification (Badapalli et al. 2023). The green wall is also playing an important role to reduce the desertification. For instance, it was clear that assessments were subjective; they could not be readily replicated, and they always relied on administrative areas rather than being taken and displayed in a continuous space. This research is trying to fulfill the mentioned research gap with the help of the existing literatures related to this field.

Identification of novel human CC chemokine receptor 8 (CCR8) antagonists via the synthesis of naphthalene amide and sulfonamide isosteres

The human CC chemokine receptor 8 (CCR8) has been extensively pursued as target for the treatment of various inflammatory disorders. More recently, the importance of CCR8 in the tumor microenvironment has been demonstrated, spurring the interest in CCR8 antagonism as therapeutic strategy in immuno-oncology. On a previously described naphthalene sulfonamide with CCR8 antagonistic properties, the concept of isosterism was applied, leading to the discovery of novel CCR8 antagonists with IC50 values in the nM range in both the CCL1 competition binding and CCR8 calcium mobilization assay. The excellent CCR8 antagonistic activity of the most potent congeners was rationalized by homology molecular modeling.

Unveiling the structural mechanisms of nonpeptide ligand recognition and activation in human chemokine receptor CCR8.

The human CC chemokine receptor 8 (CCR8) is an emerging therapeutic target for cancer immunotherapy and autoimmune diseases. Understanding the molecular recognition of CCR8, particularly with nonpeptide ligands, is valuable for drug development. Here, we report three cryo-electron microscopy structures of human CCR8 complexed with Gi trimers in the ligand-free state or activated by nonpeptide agonists LMD-009 and ZK 756326. A conserved Y1.39Y3.32E7.39 motif in the orthosteric binding pocket is shown to play a crucial role in the chemokine and nonpeptide ligand recognition. Structural and functional analyses indicate that the lack of conservation in Y1143.33 and Y1724.64 among the CC chemokine receptors could potentially contribute to the selectivity of the nonpeptide ligand binding to CCR8. These findings present the characterization of the molecular interaction between a nonpeptide agonist and a chemokine receptor, aiding the development of therapeutics targeting related diseases through a structure-based approach.

Structural basis of antibody inhibition and chemokine activation of the human CC chemokine receptor 8

Structural basis of antibody inhibition and chemokine activation of the human CC chemokine receptor 8

Development of a cellular model to study CCR8 signaling in tumor-infiltrating regulatory T cells

The human CC chemokine receptor 8 (CCR8) is specifically expressed on tumor-infiltrating regulatory T cells (TITRs) and is a promising drug target for cancer immunotherapy. However, the role of CCR8 signaling in TITR biology and the effectiveness of CCR8 small molecule antagonists as TITR-targeting immunotherapy remain subjects of ongoing debate. In this work, we generated a novel cellular model of TITRs by culturing peripheral blood mononuclear cell-derived regulatory T cells in medium containing tumor cell-conditioned medium, CD3/CD28 activator, interleukin-2 and 1α,25-dihydroxyvitamin D3. This cellular model (named TITR mimics) highly and stably expressed a series of TITR signature molecules, including CCR8, FOXP3, CD30, CD39, CD134, CD137, TIGIT and Tim-3. Moreover, TITR mimics displayed robust in vitro immunosuppressive activity. To unravel the functional role of CCR8 in TITR mimics, a chemotaxis assay was performed showing strong and CCR8-specific migration toward CCL1, the natural chemokine agonist of CCR8. However, either stimulation (with CCL1) or blocking (with the small molecule antagonist NS-15) of CCR8 signaling did not affect the immunosuppressive activity, proliferation and survival of TITR mimics. Collectively, our work provides a method for the generation of TITR mimics in vitro, which can be used to study TITR biology and to evaluate drug candidates targeting TITRs. Furthermore, our findings suggest that CCR8 signaling primarily regulates migration of these cells.

Structural basis of antibody inhibition and chemokine activation of the human CC chemokine receptor 8

The C-C motif chemokine receptor 8 (CCR8) is a class A G-protein coupled receptor that has emerged as a promising therapeutic target in cancer. Targeting CCR8 with an antibody has appeared to be an attractive therapeutic approach, but the molecular basis for chemokine-mediated activation and antibody-mediated inhibition of CCR8 are not fully elucidated. Here, we obtain an antagonist antibody against human CCR8 and determine structures of CCR8 in complex with either the antibody or the endogenous agonist ligand CCL1. Our studies reveal characteristic antibody features allowing recognition of the CCR8 extracellular loops and CCL1-CCR8 interaction modes that are distinct from other chemokine receptor - ligand pairs. Informed by these structural insights, we demonstrate that CCL1 follows a two-step, two-site binding sequence to CCR8 and that antibody-mediated inhibition of CCL1 signaling can occur by preventing the second binding event. Together, our results provide a detailed structural and mechanistic framework of CCR8 activation and inhibition that expands our molecular understanding of chemokine - receptor interactions and offers insight into the development of therapeutic antibodies targeting chemokine GPCRs.

A key GPCR phosphorylation motif discovered in arrestin2⋅CCR5 phosphopeptide complexes

The two non-visual arrestins, arrestin2 and arrestin3, bind hundreds of GPCRs with different phosphorylation patterns, leading to distinct functional outcomes. Structural information on these interactions is available only for very few GPCRs. Here, we have characterized the interactions between the phosphorylated human CC chemokine receptor 5 (CCR5) and arrestin2. We identified several new CCR5 phosphorylation sites necessary for stable arrestin2 complex formation. Structures of arrestin2 in the apo form and complexes with CCR5 C-terminal phosphopeptides, together with NMR, biochemical, and functional assays, revealed three phosphoresidues in a pXpp motif that are essential for arrestin2 binding and activation. The identified motif appears responsible for robust arrestin2 recruitment in many other GPCRs. An analysis of receptor sequences and available structural and functional information provides hints on the molecular basis of arrestin2/arrestin3 isoform specificity. Our findings demonstrate how multi-site phosphorylation controls GPCR⋅arrestin interactions and provide a framework to probe the intricate details of arrestin signaling.

Optimization of triazolo[4,5-d]pyrimidines towards human CC chemokine receptor 7 (CCR7) antagonists.

CCR7 signaling directs the migration of both immune cells and cancer cells to the lymph nodes, is involved in numerous chronic inflammatory disorders and lymph node metastases. Despite the therapeutic promise of CCR7 antagonists, no potent and selective small molecule CCR7 antagonists have been reported to date. Since most human chemokine G protein-coupled receptors (GPCRs) share a conserved intracellular allosteric binding site, new CCR7 antagonist chemotypes may be identified by screening small molecules that are known to target this site in other chemokine GPCRs. In this work, our previously prepared series of 14 scaffold-modified analogues of a known thiazolo[4,5-d]pyrimidine CXCR2 antagonist were screened as potential CCR7 antagonists. This resulted in the discovery of a triazolo[4,5-d]pyrimidine analogue with an IC50 of 2.43μM against CCR7 and 0.66μM against CXCR2. Exploration of the structure-activity relationship (SAR) for the 3-, 5- and 7-position substituents of this triazolo[4,5-d]pyrimidine resulted in improved potency and selectivity, with an IC50 of 0.43μM and 11.02μM against CCR7 and CXCR2, respectively, for the most selective derivative. Molecular docking showed that the binding mode of these triazolo[4,5-d]pyrimidines in CCR7 and CXCR2 corresponds with those of previously co-crystallized ligands.

Myeloid Progenitor Inhibitory Factor-1 (CCL23) Inhibits Lung Leukocyte Recruitment in a Primate Cardiopulmonary Bypass-Induced Pulmonary Ischaemia Model.

Bone marrow (BM)-derived polymorphonuclear leukocytes (PMNs) and monocytes (MO) induced by cardiopulmonary bypass (CPB) are highly proteolytic and cause postoperative lung injury. Although CCL23/Myeloid progenitor inhibitory factor-1 is a human CC chemokine with potent suppressor effects on myeloid progenitor cells, invivo inhibitory effects on BM-derived leukocyte kinetics associated with CPB are unknown. Two-hour CPB was surgically performed in cynomolgus monkeys and BM-derived leukocytes kinetics were monitored postoperatively by flow cytometry with 5'-bromo-2'-deoxyuridine (BrdU) and cytokine ELISA. Monkeys were given CCL23 (n=5) or saline (control, n=5) intravenously daily for 3 days before BrdU labelling and peripheral blood/bronchoalveolar lavage fluid (BALF) timepoint sampling to reveal BrdU-labelled cells. Levels of cytokines, CD11b, and L-selectin were considered leukocytic activation markers. The CCL23 treatment significantly prolonged BM transit of leukocytes (PMNs, 118.4±11.7-95.5±4.1 hours [control]; MO, 91.6±5.0-62.0±3.0 hours [control]) and reduced their alveolar appearance. The BM pool size of MO was decreased by CCL23 but PMNs were unaffected. CD11b, L-selectin expression of PMNs and MO during CPB, and post-surgical increases of interleukin (IL)-6, IL-8, TNF-α, MCP-1, and PMN elastase in the BALF were not suppressed. CCL23 treatment slows turnover of PMN and MO progenitors in BM and suppresses their circulatory release and lung recruitment. CCL23 has inhibitory effects specifically on the CPB-induced BM response and could hold value for preventing CPB-induced lung injury.

Courses of Arginine-Vasopressin in the Systemic and Cavernous Blood through Different Stages of Sexual Arousal in Healthy Males and Patients with Erectile Dysfunction

To investigate the role of the peptide arginine-vasopressin (AVP) in controlling the function of penile erectile tissue, we determined the course of AVP through different stages of sexual arousal in both the systemic and cavernous blood of healthy males and patients presenting with ED. Twenty-five healthy males and 45 patients with ED were exposed to erotic stimulation to induce sexual arousal. Blood was withdrawn from the corpus cavernosum and a cubital vein during penile flaccidity, tumescence, rigid erection (attained only by the healthy individuals), and detumescence. AVP (ng/l plasma) was determined by means of a radioimmunoassay. Effects of AVP (0.1 to 100 nM) on isolated human CC were examined using a tissue bath system. AVP elicited contraction of isolated CC. In the healthy subjects, a decline in AVP levels (5.4 to 3 ng/l) was seen in the systemic blood when the flaccid penis became rigid. In the cavernous blood, no alterations were registered. In the group of ED patients, AVP in the systemic circulation did not display a transient decline. The drop in systemic AVP in healthy males during sexual stimulation might be a prerequisite to enable penile erection.

“Glyco-sulfo barcodes” regulate chemokine receptor function

Chemokine ligands and receptors regulate the directional migration of leukocytes. Post-translational modifications of chemokine receptors including O-glycosylation and tyrosine sulfation have been reported to regulate ligand binding and resulting signaling. Through in silico analyses, we determined potential conserved O-glycosylation and sulfation sites on human and murine CC chemokine receptors. Glyco-engineered CHO cell lines were used to measure the impact of O-glycosylation on CC chemokine receptor CCR5, while mutation of tyrosine residues and treatment with sodium chlorate were performed to determine the effect of tyrosine sulfation. Changing the glycosylation or tyrosine sulfation on CCR5 reduced the receptor signaling by the more positively charged CCL5 and CCL8 more profoundly compared to the less charged CCL3. The loss of negatively charged sialic acids resulted only in a minor effect on CCL3-induced signal transduction. The enzymes GalNAc-T1 and GalNAc-T11 were shown to be involved in the process of chemokine receptor O-glycosylation. These results indicate that O-glycosylation and tyrosine sulfation are involved in the fine-tuning and recognition of chemokine interactions with CCR5 and the resulting signaling.

Molecular insights into ligand recognition and activation of chemokine receptors CCR2 and CCR3

Chemokine receptors are a family of G-protein-coupled receptors with key roles in leukocyte migration and inflammatory responses. Here, we present cryo-electron microscopy structures of two human CC chemokine receptor–G-protein complexes: CCR2 bound to its endogenous ligand CCL2, and CCR3 in the apo state. The structure of the CCL2–CCR2–G-protein complex reveals that CCL2 inserts deeply into the extracellular half of the transmembrane domain, and forms substantial interactions with the receptor through the most N-terminal glutamine. Extensive hydrophobic and polar interactions are present between both two chemokine receptors and the Gα-protein, contributing to the constitutive activity of these receptors. Notably, complemented with functional experiments, the interactions around intracellular loop 2 of the receptors are found to be conserved and play a more critical role in G-protein activation than those around intracellular loop 3. Together, our findings provide structural insights into chemokine recognition and receptor activation, shedding lights on drug design targeting chemokine receptors.

MP38-02 IMPAIRED NICOTINAMIDE ADENINE DINUCLEOTIDE (NAD+) MEDIATING ENDOTHELIUM-INDEPENDENT RELAXATION WITH AGING ON HUMAN CORPUS CAVERNOSUM

You have accessJournal of UrologyCME1 May 2022MP38-02 IMPAIRED NICOTINAMIDE ADENINE DINUCLEOTIDE (NAD+) MEDIATING ENDOTHELIUM-INDEPENDENT RELAXATION WITH AGING ON HUMAN CORPUS CAVERNOSUM Serap Gur, Suresh C. Sikka, Sudha Talwar, Didem Yilmaz-Oral, Asim Abdel-Mageed, and Wayne J. G. Hellstrom Serap GurSerap Gur More articles by this author , Suresh C. SikkaSuresh C. Sikka More articles by this author , Sudha TalwarSudha Talwar More articles by this author , Didem Yilmaz-OralDidem Yilmaz-Oral More articles by this author , Asim Abdel-MageedAsim Abdel-Mageed More articles by this author , and Wayne J. G. HellstromWayne J. G. Hellstrom More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002592.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Nicotinamide adenine dinucleotide (NAD+), a small-molecule cofactor for NAD+-dependent enzymes, such as the sirtuins (SIRT) is a coenzyme for redox reactions, making it central to energy metabolism. SIRT-stimulation by hydrogen sulfide (H2S) reverses redox changes. Remarkably, ageing is accompanied by a gradual decline in tissue and cellular NAD+ levels leading to a slow rate of glycolysis, mitochondrial electron transport, and ATP formation. This study aimed to evaluate the effect of aging on NAD+-induced relaxation of human CC. METHODS: Human CC from ED patients (n=12) undergoing penile prosthesis surgery were processed for organ bath. After precontraction with phenylephrine (Phe, 10µM), concentration-response curves were performed for NAD+ (10nM-100µM) on CC from middle-aged and aged individuals. Underlying mechanisms of relaxation were evaluated by inhibitory agents, namely L-NAME [an inhibitor of NOS], ODQ (a soluble guanylyl cyclase inhibitor), and tetraethylammonium (TEA, non-selective K+ channel blocker). The relaxation responses to electrical field stimulation (EFS), acetylcholine (ACh), sodium hydrogen sulfur (NaHS, a donor of H2S), adenosine (Ado), and contractile response to EFS were examined in the presence or absence of NAD+(10μM). Cellular localization of eNOS, nNOS, and VEGF expressions was performed by Western blotting and immunofluorescence. For the primary cell culture of human penile tissue samples, the cells in growth media for 2 weeks were characterized for smooth muscle cell protein marker, treated with 500µM of NAD+ either 6 hours or 24 hours, and cell lysate was quantitated for eNOS, nNOS, VEGF, and GAPDH expressions. RESULTS: NAD+ reduced the maximal contractile response induced by Phe (46.2±9.9%). The NAD+-induced relaxations were not affected by inhibitors. NAD+ induced relaxations were significantly lower (11.1±5.3%, p=0.0270) in the aged (75.6±1.2 years) group when compared (40.7±6.9%) middle-aged (63.0±4.0 years) group. NAD+-evoked relaxations of HCC were augmented with NaHS and Ado incubations. Using Western blot analysis, confocal imaging, and cell-culture experiments for eNOS, nNOS, and VEGF, NAD+ could not change impairment in samples. CONCLUSIONS: Our studies have elucidated a decreased relaxation of NAD+ on HCC tissues from aged men. The relaxation response to NAD+ is independent of the activation of the NO-cGMP system. NAD+-involved molecular targets can be through actions at H2S and Ado- receptor mechanisms in HCC. Collectively, NAD+ repletion as a promising therapeutic approach may preserve penile biogenesis and integrity during aging. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e618 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Serap Gur More articles by this author Suresh C. Sikka More articles by this author Sudha Talwar More articles by this author Didem Yilmaz-Oral More articles by this author Asim Abdel-Mageed More articles by this author Wayne J. G. Hellstrom More articles by this author Expand All Advertisement PDF DownloadLoading ...

A Set of Experimentally Validated Decoys for the Human CC Chemokine Receptor 7 (CCR7) Obtained by Virtual Screening.

We present a state-of-the-art virtual screening workflow aiming at the identification of novel CC chemokine receptor 7 (CCR7) antagonists. Although CCR7 is associated with a variety of human diseases, such as immunological disorders, inflammatory diseases, and cancer, this target is underexplored in drug discovery and there are no potent and selective CCR7 small molecule antagonists available today. Therefore, computer-aided ligand-based, structure-based, and joint virtual screening campaigns were performed. Hits from these virtual screenings were tested in a CCL19-induced calcium signaling assay. After careful evaluation, none of the in silico hits were confirmed to have an antagonistic effect on CCR7. Hence, we report here a valuable set of 287 inactive compounds that can be used as experimentally validated decoys.

Structure-guided engineering of tick evasins for targeting chemokines in inflammatory diseases

As natural chemokine inhibitors, evasin proteins produced in tick saliva are potential therapeutic agents for numerous inflammatory diseases. Engineering evasins to block the desired chemokines and avoid off-target side effects requires structural understanding of their target selectivity. Structures of the class A evasin EVA-P974 bound to human CC chemokine ligands 7 and 17 (CCL7 and CCL17) and to a CCL8-CCL7 chimera reveal that the specificity of class A evasins for chemokines of the CC subfamily is defined by conserved, rigid backbone-backbone interactions, whereas the preference for a subset of CC chemokines is controlled by side-chain interactions at four hotspots in flexible structural elements. Hotspot mutations alter target preference, enabling inhibition of selected chemokines. The structure of an engineered EVA-P974 bound to CCL2 reveals an underlying molecular mechanism of EVA-P974 target preference. These results provide a structure-based framework for engineering evasins as targeted antiinflammatory therapeutics.

A Comprehensive Computational Screening of Phytochemicals Derived from Saudi Medicinal Plants against Human CC Chemokine Receptor 7 to Identify Potential Anti-Cancer Therapeutics.

Homeostatic trafficking of immune cells by CC chemokine receptor 7 (CCR7) keeps immune responses and tolerance in a balance. The involvement of this protein in lymph node metastasis in cancer marks CCR7 as a penitential drug target. Using the crystal structure of CCR7, herein, a comprehensive virtual screening study is presented to filter novel strong CCR7 binding phytochemicals from Saudi medicinal plants that have a higher binding affinity for the intracellular allosteric binding pocket. By doing so, three small natural molecules named as Hit-1 (1,8,10-trihydroxy-3-methoxy-6-methylanthracen-9(4H)-one), Hit-2 (4-(3,4-dimethoxybenzyl)-3-(4-hydroxy-3-methoxybenzyl)dihydrofuran-2(3H)-one), and Hit-3 (10-methyl-12,13-dihydro-[1,2]dioxolo[3,4,5-de]furo[3,2-g]isochromeno[4,3-b]chromen-8-ol) are predicted showing strong binding potential for the CC chemokine receptor 7 allosteric pocket. During molecular dynamics simulations, the compounds were observed in the formation of several chemical bonding of short bond distances. Additionally, the molecules remained in strong contact with the active pocket residues and experienced small conformation changes that seemed to be mediated by the CCR7 loops to properly engage the ligands. Two types of binding energy methods (MM/GBPBSA and WaterSwap) were additionally applied to further validate docking and simulation findings. Both analyses complement the good affinity of compounds for CCR7, the electrostatic and van der Waals energies being the most dominant in intermolecular interactions. The active pocket residue’s role in compounds binding was further evaluated via alanine scanning, which highlighted their importance in natural compounds binding. Additionally, the compounds fulfilled all drug-like rules: Lipinski, Ghose, Veber, Egan, and Muegge passed many safety parameters, making them excellent anti-cancer candidates for experimental testing.

Structural basis of the activation of the CC chemokine receptor 5 by a chemokine agonist.

The human CC chemokine receptor 5 (CCR5) is a G protein-coupled receptor (GPCR) that plays a major role in inflammation and is involved in cancer, HIV, and COVID-19. Despite its importance as a drug target, the molecular activation mechanism of CCR5, i.e., how chemokine agonists transduce the activation signal through the receptor, is yet unknown. Here, we report the cryo-EM structure of wild-type CCR5 in an active conformation bound to the chemokine super-agonist [6P4]CCL5 and the heterotrimeric Gi protein. The structure provides the rationale for the sequence-activity relation of agonist and antagonist chemokines. The N terminus of agonist chemokines pushes onto specific structural motifs at the bottom of the orthosteric pocket that activate the canonical GPCR microswitch network. This activation mechanism differs substantially from other CC chemokine receptors that bind chemokines with shorter N termini in a shallow binding mode involving unique sequence signatures and a specialized activation mechanism.