Education; Immune System Diseases; Inflammation; Neoplasms; Parasitic Diseases; Pharmacology; Crystallography, X-Ray; Enzymes and Coenzymes; High-Throughput Screening Assays
We are interested in understanding the mechanism of action of proteins involved in infectious disease, inflammation, and cancer using a variety of techniques including structural biology, yeast genetics, and signal transduction. For example, we determined the three-dimensional structure of two chemokines, human CXCL12 and herpesvirus-8 vMIP-II. We are currently using variety of methods to determine how these proteins interact with theG-protein coupled receptor, CXCR4, a HIV-1 co-receptor and a therapeutic target for cancer metastasis. In collaboration with Demetrios Braddock (Departmentof Pathology), we determined the structure of FIR, a protein that counter-regulates the transcription of the oncogene c-myc. In the area of inflammation we have been studying the signaling mechanism by which macrophage migration inhibitory factor (MIF) exerts its pro-inflammatory effects. We have determined the structure of MIF in the presence of small molecule inhibitors and are studying these inhibitors in mouse models of autoimmunity, inflammation, or infectious diseases.
Specialized Terms: Cancer; Inflammation; Infectious disease; Macromolecular X-ray crystallography; Signaling pathways; Drug design; High throughput screening (HTPS)
Extensive Research Description
We are generally interested in understanding the biology, structure, mechanism of action, and pharmacology (inhibition) of proteins that lead to infectious disease, inflammation, and cancer. Our studies are multidisciplinary and include structural biology (X-ray crystallography or NMR), high throughput screening and/or inhibitor design, structure-activity relationships, mutational analysis, the use of a strain of yeast as a model system for chemokines and their receptors, and animal models of disease.
Our current studies include structure-function relationships of chemokines, a family of secreted proteins that recruit immune cells to the site of infection or tissue injury and function by activating specific G-protein coupled receptors. CXCL12 (SDF-1alpha) and vMIP-II/vCCL2 are an agonist and antagonist, respectively, for the GPCR CXCR4, a co-receptor for HIV-1. This receptor also mediates metastasis in a number of cancers, and a C-terminal mutation responsible for an immunosuppressive disease known as WHIM syndrome. Importantly, CXCR4 is the target for Plerixafor, a drug used in peripheral stem cell transplantation. We have determined the structures of CXCL12 and vMIP-II/vCCL2 and are working towards understanding their interactions CXCR4 at high resolution. We are also using random mutagenesis combined with yeast genetics to define smaller peptides with agonist, partial agonist, or antagonist activity.
The other major project is MIF. Inhibition of this "cytokine" MIF by neutralizing antibodies leads to therapeutic efficacy in animal models of sepsis, arthritis, and glomerulonephritis. This protein also inhibits the apoptotic activity of p53. The three-dimensional structure of MIF revealed a striking similarity with two microbial enzymes, including the presence of an enzyme-like crevice with catalytic functional groups. We are now attempting to determine the role of this site in inflammatory diseases and cancer. Interestingly, parasites also express and secrete this protein. In collaborative studies with Drs. Michael Cappello and Richard Bucala, we have determined the structure of MIF expressed from hookworm and Leishmania major, used high throughput screening, and structure-activity relationships to identify compounds with sub-micromolar Ki's.
Other projects include determining the structure of (1) signal transduction proteins involved in inflammation, and (2) enzymes that degrade chemotherapeutic agents.
1. Chemokine-chemokine receptor (GPCR) structures
2. High throughput screening to identify small molecule agonists and antagonists of chemokines and their GPCR receptors
3. Yeast genetics to identify and quantitate chemokine-chemokine receptor interactions
4. Mechanism of receptor activation for macrophage migration inhibitory factor
5. Identification of the substrate of MIF and structure of MIF-ligand complexes
6. Study of MIF inhibitors in disease models
7. Co-crystal structures of other protein-inhibitors complexes
- Macrophage Migration Inhibitory Factor-CXCR4 Receptor Interactions: EVIDENCE FOR PARTIAL ALLOSTERIC AGONISM IN COMPARISON WITH CXCL12 CHEMOKINE. J Biol Chem. 2016 Jul 22;291(30):15881-95. doi: 10.1074/jbc.M116.717751.
- An Analysis of MIF Structural Features that Control Functional Activation of CD74. Chem Biol. 2015 Sep 17;22(9):1197-205. doi: 10.1016/j.chembiol.2015.08.006.
- Crystallographic and receptor binding characterization of Plasmodium falciparum macrophage migration inhibitory factor complexed to two potent inhibitors. J Med Chem. 2014 Oct 23;57(20):8652-6. doi: 10.1021/jm501168q.
- Targeting distinct tautomerase sites of D-DT and MIF with a single molecule for inhibition of neutrophil lung recruitment. FASEB J. 2014 Nov;28(11):4961-71. doi: 10.1096/fj.14-256636.
- Structural insight into the evolution of a new chemokine family from zebrafish. Proteins. 2014 May;82(5):708-16. doi: 10.1002/prot.24380.
- MIF intersubunit disulfide mutant antagonist supports activation of CD74 by endogenous MIF trimer at physiologic concentrations. Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10994-9. doi: 10.1073/pnas.1221817110.
- Allosteric peptide regulators of chemokine receptors CXCR4 and CXCR7. Biochem Pharmacol. 2013 Nov 1;86(9):1263-71. doi: 10.1016/j.bcp.2013.08.019. Epub 2013 Aug 22.
Full List of PubMed Publications
- Gil-Yarom N, Radomir L, Sever L, Kramer MP, Lewinsky H, Bornstein C, Blecher-Gonen R, Barnett-Itzhaki Z, Mirkin V, Friedlander G, Shvidel L, Herishanu Y, Lolis EJ, Becker-Herman S, Amit I, Shachar I: CD74 is a novel transcription regulator. Proc Natl Acad Sci U S A. 2017 Jan 17; 2016 Dec 28. PMID: 28031488
- Meza-Romero R, Benedek G, Jordan K, Leng L, Pantouris G, Lolis E, Bucala R, Vandenbark AA: Modeling of both shared and distinct interactions between MIF and its homologue D-DT with their common receptor CD74. Cytokine. 2016 Dec; 2016 Aug 27. PMID: 27573366
- Rajasekaran D, Gröning S, Schmitz C, Zierow S, Drucker N, Bakou M, Kohl K, Mertens A, Lue H, Weber C, Xiao A, Luker G, Kapurniotu A, Lolis E, Bernhagen J: Macrophage Migration Inhibitory Factor-CXCR4 Receptor Interactions: EVIDENCE FOR PARTIAL ALLOSTERIC AGONISM IN COMPARISON WITH CXCL12 CHEMOKINE. J Biol Chem. 2016 Jul 22; 2016 May 19. PMID: 27226569
- Yang Y, Hodsdon ME, Lolis EJ, Ehrlich BE: Conformational dynamics of Ca2+-dependent responses in the polycystin-2 C-terminal tail. Biochem J. 2016 Feb 1; 2015 Nov 16. PMID: 26574436
- Stoppe C, Rex S, Goetzenich A, Kraemer S, Emontzpohl C, Soppert J, Averdunk L, Sun Y, Rossaint R, Lue H, Huang C, Song Y, Pantouris G, Lolis E, Leng L, Schulte W, Bucala R, Weber C, Bernhagen J: Interaction of MIF Family Proteins in Myocardial Ischemia/Reperfusion Damage and Their Influence on Clinical Outcome of Cardiac Surgery Patients. Antioxid Redox Signal. 2015 Oct 10; 2015 Aug 11. PMID: 26234719
- Pantouris G, Syed MA, Fan C, Rajasekaran D, Cho TY, Rosenberg EM Jr, Bucala R, Bhandari V, Lolis EJ: An Analysis of MIF Structural Features that Control Functional Activation of CD74. Chem Biol. 2015 Sep 17; 2015 Sep 10. PMID: 26364929
- Yang Y, Keeler C, Kuo IY, Lolis EJ, Ehrlich BE, Hodsdon ME: Oligomerization of the polycystin-2 C-terminal tail and effects on its Ca2+-binding properties. J Biol Chem. 2015 Apr 17; 2015 Feb 25. PMID: 25716316
- Rajasekaran D, Zierow S, Syed M, Bucala R, Bhandari V, Lolis EJ: Targeting distinct tautomerase sites of D-DT and MIF with a single molecule for inhibition of neutrophil lung recruitment. FASEB J. 2014 Nov; 2014 Jul 11. PMID: 25016026
- Pantouris G, Rajasekaran D, Garcia AB, Ruiz VG, Leng L, Jorgensen WL, Bucala R, Lolis EJ: Crystallographic and receptor binding characterization of Plasmodium falciparum macrophage migration inhibitory factor complexed to two potent inhibitors. J Med Chem. 2014 Oct 23; 2014 Oct 8. PMID: 25268646
- Ioannou K, Cheng KF, Crichlow GV, Birmpilis AI, Lolis EJ, Tsitsilonis OE, Al-Abed Y: ISO-66, a novel inhibitor of macrophage migration, shows efficacy in melanoma and colon cancer models. Int J Oncol. 2014 Oct; 2014 Jul 22. PMID: 25050663
- Rajasekaran D, Fan C, Meng W, Pflugrath JW, Lolis EJ: Structural insight into the evolution of a new chemokine family from zebrafish. Proteins. 2014 May; 2013 Dec 14. PMID: 23900850
- Fan C, Rajasekaran D, Syed MA, Leng L, Loria JP, Bhandari V, Bucala R, Lolis EJ: MIF intersubunit disulfide mutant antagonist supports activation of CD74 by endogenous MIF trimer at physiologic concentrations. Proc Natl Acad Sci U S A. 2013 Jul 2; 2013 Jun 17. PMID: 23776208
- Crichlow GV, Fan C, Keeler C, Hodsdon M, Lolis EJ: Structural interactions dictate the kinetics of macrophage migration inhibitory factor inhibition by different cancer-preventive isothiocyanates. Biochemistry. 2012 Sep 25; 2012 Sep 11. PMID: 22931430
- Sun T, Holowka T, Song Y, Zierow S, Leng L, Chen Y, Xiong H, Griffith J, Nouraie M, Thuma PE, Lolis E, Janse CJ, Gordeuk VR, Augustijn K, Bucala R: A Plasmodium-encoded cytokine suppresses T-cell immunity during malaria. Proc Natl Acad Sci U S A. 2012 Jul 31; 2012 Jul 9. PMID: 22778413
- Rajasekaran D, Keeler C, Syed MA, Jones MC, Harrison JK, Wu D, Bhandari V, Hodsdon ME, Lolis EJ: A model of GAG/MIP-2/CXCR2 interfaces and its functional effects. Biochemistry. 2012 Jul 17; 2012 Jul 2. PMID: 22686371
- Cho Y, Vermeire JJ, Merkel JS, Leng L, Du X, Bucala R, Cappello M, Lolis E: Drug repositioning and pharmacophore identification in the discovery of hookworm MIF inhibitors. Chem Biol. 2011 Sep 23. PMID: 21944748
- Merk M, Zierow S, Leng L, Das R, Du X, Schulte W, Fan J, Lue H, Chen Y, Xiong H, Chagnon F, Bernhagen J, Lolis E, Mor G, Lesur O, Bucala R: The D-dopachrome tautomerase (DDT) gene product is a cytokine and functional homolog of macrophage migration inhibitory factor (MIF). Proc Natl Acad Sci U S A. 2011 Aug 23; 2011 Aug 4. PMID: 21817065
- Choi M, Scholl UI, Yue P, Björklund P, Zhao B, Nelson-Williams C, Ji W, Cho Y, Patel A, Men CJ, Lolis E, Wisgerhof MV, Geller DS, Mane S, Hellman P, Westin G, Åkerström G, Wang W, Carling T, Lifton RP: K+ channel mutations in adrenal aldosterone-producing adenomas and hereditary hypertension. Science. 2011 Feb 11. PMID: 21311022
- Leng L, Chen L, Fan J, Greven D, Arjona A, Du X, Austin D, Kashgarian M, Yin Z, Huang XR, Lan HY, Lolis E, Nikolic-Paterson D, Bucala R: A small-molecule macrophage migration inhibitory factor antagonist protects against glomerulonephritis in lupus-prone NZB/NZW F1 and MRL/lpr mice. J Immunol. 2011 Jan 1; 2010 Nov 24. PMID: 21106847
- Kulkarni MV, Tettamanzi MC, Murphy JW, Keeler C, Myszka DG, Chayen NE, Lolis EJ, Hodsdon ME: Two independent histidines, one in human prolactin and one in its receptor, are critical for pH-dependent receptor recognition and activation. J Biol Chem. 2010 Dec 3; 2010 Sep 30. PMID: 20889499
- Cho Y, Crichlow GV, Vermeire JJ, Leng L, Du X, Hodsdon ME, Bucala R, Cappello M, Gross M, Gaeta F, Johnson K, Lolis EJ: Allosteric inhibition of macrophage migration inhibitory factor revealed by ibudilast. Proc Natl Acad Sci U S A. 2010 Jun 22; 2010 Jun 8. PMID: 20534506
- Murphy JW, Yuan H, Kong Y, Xiong Y, Lolis EJ: Heterologous quaternary structure of CXCL12 and its relationship to the CC chemokine family. Proteins. 2010 Apr. PMID: 20077567
- Crichlow GV, Lubetsky JB, Leng L, Bucala R, Lolis EJ: Structural and kinetic analyses of macrophage migration inhibitory factor active site interactions. Biochemistry. 2009 Jan 13. PMID: 19090677
- Vermeire JJ, Cho Y, Lolis E, Bucala R, Cappello M: Orthologs of macrophage migration inhibitory factor from parasitic nematodes. Trends Parasitol. 2008 Aug; 2008 Jul 4. PMID: 18603473
- Kamir D, Zierow S, Leng L, Cho Y, Diaz Y, Griffith J, McDonald C, Merk M, Mitchell RA, Trent J, Chen Y, Kwong YK, Xiong H, Vermeire J, Cappello M, McMahon-Pratt D, Walker J, Bernhagen J, Lolis E, Bucala R: A Leishmania ortholog of macrophage migration inhibitory factor modulates host macrophage responses. J Immunol. 2008 Jun 15. PMID: 18523291
- Zhang W, Cho Y, Lolis E, Howe JR: Structural and single-channel results indicate that the rates of ligand binding domain closing and opening directly impact AMPA receptor gating. J Neurosci. 2008 Jan 23. PMID: 18216201
- Crichlow GV, Zhou H, Hsiao HH, Frederick KB, Debrosse M, Yang Y, Folta-Stogniew EJ, Chung HJ, Fan C, De la Cruz EM, Levens D, Lolis E, Braddock D: Dimerization of FIR upon FUSE DNA binding suggests a mechanism of c-myc inhibition. EMBO J. 2008 Jan 9; 2007 Dec 6. PMID: 18059478
- Cho Y, Jones BF, Vermeire JJ, Leng L, DiFedele L, Harrison LM, Xiong H, Kwong YK, Chen Y, Bucala R, Lolis E, Cappello M: Structural and functional characterization of a secreted hookworm Macrophage Migration Inhibitory Factor (MIF) that interacts with the human MIF receptor CD74. J Biol Chem. 2007 Aug 10; 2007 Jun 13. PMID: 17567581
- Crichlow GV, Cheng KF, Dabideen D, Ochani M, Aljabari B, Pavlov VA, Miller EJ, Lolis E, Al-Abed Y: Alternative chemical modifications reverse the binding orientation of a pharmacophore scaffold in the active site of macrophage migration inhibitory factor. J Biol Chem. 2007 Aug 10; 2007 May 25. PMID: 17526494
- Murphy JW, Cho Y, Sachpatzidis A, Fan C, Hodsdon ME, Lolis E: Structural and functional basis of CXCL12 (stromal cell-derived factor-1 alpha) binding to heparin. J Biol Chem. 2007 Mar 30; 2007 Jan 29. PMID: 17264079
- Shi X, Leng L, Wang T, Wang W, Du X, Li J, McDonald C, Chen Z, Murphy JW, Lolis E, Noble P, Knudson W, Bucala R: CD44 is the signaling component of the macrophage migration inhibitory factor-CD74 receptor complex. Immunity. 2006 Oct. PMID: 17045821
- Bucala R, Lolis E: Macrophage migration inhibitory factor: a critical component of autoimmune inflammatory diseases. Drug News Perspect. 2005 Sep. PMID: 16362080
- Scheuermann TH, Lolis E, Hodsdon ME: Tertiary structure of thiopurine methyltransferase from Pseudomonas syringae, a bacterial orthologue of a polymorphic, drug-metabolizing enzyme. J Mol Biol. 2003 Oct 24. PMID: 14556746
- Lolis E, Bucala R: Therapeutic approaches to innate immunity: severe sepsis and septic shock. Nat Rev Drug Discov. 2003 Aug. PMID: 12904813
- Lolis E, Bucala R: Macrophage migration inhibitory factor. Expert Opin Ther Targets. 2003 Apr. PMID: 12667094
- Sachpatzidis A, Benton BK, Manfredi JP, Wang H, Hamilton A, Dohlman HG, Lolis E: Identification of allosteric peptide agonists of CXCR4. J Biol Chem. 2003 Jan 10; 2002 Nov 3. PMID: 12417595
- Lubetsky JB, Dios A, Han J, Aljabari B, Ruzsicska B, Mitchell R, Lolis E, Al-Abed Y: The tautomerase active site of macrophage migration inhibitory factor is a potential target for discovery of novel anti-inflammatory agents. J Biol Chem. 2002 Jul 12; 2002 May 7. PMID: 11997397
- Sankoorikal BJ, Zhu YL, Hodsdon ME, Lolis E, Dannies PS: Aggregation of human wild-type and H27A-prolactin in cells and in solution: roles of Zn(2+), Cu(2+), and pH. Endocrinology. 2002 Apr. PMID: 11897686
- Fernandez EJ, Lolis E: Structure, function, and inhibition of chemokines. Annu Rev Pharmacol Toxicol. 2002. PMID: 11807180