G protein coupled receptors (GPCRs) represent the largest protein superfamily in humans, with nearly 1000 members. They are seven transmembrane receptors that coordinate intercellular communication via the transduction of a wide range of stimuli involved in sensation, neurotransmission, development, emotion, cognition, and function in the CNS, endocrine and immune systems. Chemokine receptors are an important class of GPCRs that are best known for their pivotal role in immune surveillance, where they control the migration and activation of leukocytes in an effort to detect and resolve physiological abnormalities such as cancer and infection. However, inappropriate expression or regulation of these receptors is associated with an extraordinary number of pathologies including inflammatory diseases, cancer and AIDS;thus there is significant interest in developing small molecule receptor antagonists that block the function of specific chemokine receptors. Until recently, GPCRs had eluded structure determination due to challenges in receptor expression and crystallization. However, new technologies have emerged which has made the viability of determining GPCR structures indisputable. To this end, our primary goal is to obtain structural information on chemokine receptors and receptor complexes that can aid drug discovery efforts aimed at improving affinity, efficacy, and selectivity. Accordingly, in collaboration with the PSI network, we will apply novel technologies for the expression, purification and crystallization of GPCRs from the chemokine receptor family, with the goal of determining at least two different receptor structures and multiple co-complexes by the five-year endpoint. To maximize the capabilities of the PSI centers in generating purified protein, and to acquire insights into the dynamic aspects of receptor function, the crystallographic work will be complemented with biophysical studies. Radiolytic footprinting will be developed and applied to map the binding interfaces between chemokines and receptors and to determine information on activation mechanisms. The interaction of pathogenic proteins with chemokine receptors will also be investigated, specifically, the CCR5 receptor with the HIV glycoprotein gp120, and the DARC receptor with the malarial docking protein, DBP. Site Directed Spin Labeling with Electron Paramagnetic Resonance (SDSL-EPR) will be used to characterize the conformational changes associated with ligand binding. All of these studies will be augmented with computational modeling methods in order to rationally guide the experimental construct design and to interpret the biophysical data in a 3D context.
Chemokine receptors are involved in many human diseases including asthma, multiple sclerosis, rheumatoid arthritis, heart disease, cancer and HIV, Determining structures of chemokine receptors in complex with small molecule drugs and ligands, and information on ligand-induced conformational changes, will significantly aid the development of drugs targeting these receptors.
|Gustavsson, Martin; Zheng, Yi; Handel, Tracy M (2016) Production of Chemokine/Chemokine Receptor Complexes for Structural Biophysical Studies. Methods Enzymol 570:233-60|
|Leach, Katie; Gregory, Karen J; Kufareva, Irina et al. (2016) Towards a structural understanding of allosteric drugs at the human calcium-sensing receptor. Cell Res 26:574-92|
|Zheng, Yi; Qin, Ling; Zacarías, Natalia V Ortiz et al. (2016) Structure of CC chemokine receptor 2 with orthosteric and allosteric antagonists. Nature 540:458-461|
|Ngo, Tony; Kufareva, Irina; Coleman, James Lj et al. (2016) Identifying ligands at orphan GPCRs: current status using structure-based approaches. Br J Pharmacol 173:2934-51|
|Husby, Jarmila; Bottegoni, Giovanni; Kufareva, Irina et al. (2015) Structure-based predictions of activity cliffs. J Chem Inf Model 55:1062-76|
|Geidl, Stanislav; Svobodová Va?eková, Radka; Bendová, Veronika et al. (2015) How Does the Methodology of 3D Structure Preparation Influence the Quality of pKa Prediction? J Chem Inf Model 55:1088-97|
|Kufareva, Irina; Handel, Tracy M; Abagyan, Ruben (2015) Experiment-Guided Molecular Modeling of Protein-Protein Complexes Involving GPCRs. Methods Mol Biol 1335:295-311|
|Lenoir, Marc; Kufareva, Irina; Abagyan, Ruben et al. (2015) Membrane and Protein Interactions of the Pleckstrin Homology Domain Superfamily. Membranes (Basel) 5:646-63|
|Kufareva, Irina; Salanga, Catherina L; Handel, Tracy M (2015) Chemokine and chemokine receptor structure and interactions: implications for therapeutic strategies. Immunol Cell Biol 93:372-83|
|Qin, Ling; Kufareva, Irina; Holden, Lauren G et al. (2015) Structural biology. Crystal structure of the chemokine receptor CXCR4 in complex with a viral chemokine. Science 347:1117-22|
Showing the most recent 10 out of 51 publications