Curriculum Vitae BEILI WU The Stevens’ Lab Department of Molecular Biology The Scripps Research Institute, La Jolla, USA Telephone: 1-858-784-9411 (Lab), 1-858-366-8291 (Cell); E-mail: [email protected] PERSONAL INFORMATION First name: Beili Surname: Wu Sex: Female Date of birth: January 28th, 1979 Address: 3950 Mahaila Ave. APT. U13, San Diego, CA 92122, USA (home). The Scripps Research Institute, 10550 North Pines Road, GAC-1200, La Jolla, CA 92037, USA (Lab). EDUCATION AND TRAINING April, 2007 – present Research Associate Department of Molecular Biology, The Scripps Research Institute, La Jolla, USA September, 2001 - July, 2006 Ph.D. of Science Department of Biological Science and Biotechnology (September, 2001 – April, 2005) & Medical School (April, 2005 – July, 2006) Tsinghua University, Beijing, China September, 1997 - July, 2001 Bachelor of Science Department of Biology Beijing Normal University, Beijing, China AWARDS AND HONORS 2004 Huipu Scholarship, Tsinghua University 2002-2003 Guanghua Scholarship, Tsinghua University 2002 Lab Foundation and Contribution Scholarship, Tsinghua University 2001 Outstanding Graduate, Beijing Normal University; Outstanding Graduate, All the universities of Beijing 2000 Baogang Education Scholarship, Beijing Normal University 1999-2000 Second-Class Prize, Beijing Normal University 1998-1999 First-Class Prize, Beijing Normal University 1997-1998 Second-Class Prize, Beijing Normal University PUBLICATIONS 1. Wu B, Chien EY, Mol CD, Fenalti G, Liu W, Katritch V, Abagyan R, Brooun A, Wells P, Bi FC, Hamel DJ, Kuhn P, Handel TM, Cherezov V & Stevens RC. Structures of the CXCR4 chemokine receptor in complex with small molecule and cyclic peptide antagonists. Science. (in press) 2. Wu B, Li P, Liu Y, Lou Z, Ding Y, Shu C, Ye S, Bartlam M, Shen B & Rao Z. 3D structure of human FK506-binding protein 52: implications for the assembly of the glucocorticoid receptor/Hsp90/immunophilin heterocomplex. Proc Natl Acad Sci USA. 2004, 101:8348-8353 3. Wu B, Li P, Shu C, Shen B and Rao Z. Crystallization and preliminary crystallographic studies on the C-terminal Domain of human FKBP52. Acta Cryst. D. 2003, 59:2269-2271 4. Wu B, Liu Y, Zhao Q, Liao S, Zhang J, Bartlam M, Chen W & Rao Z. Crystal Structure of RS21-C6, involved in nucleoside triphosphate pyrophosphohydrolysis. Journal of Molecular Biology. 2007, 367 (5): 1405-12 5. Li P, Ding Y, Wu B, Shu C, Shen B and Rao Z. Crystal Structure of the N-terminal domain of human FKBP52. Acta Cryst. D. 2003, 59:16-22 6. Li P, Zhu J, Wu B, Gao F, Tien P, Rao Z and Gao GF. Crystallization and preliminary X-ray diffraction analysis of post-fusion six-helix bundle core structure from Newcastle disease virus F protein. Acta Cryst. D. 2003, 59:1296-1298 7. Li P, Wang L, Wu B, Shu C, Nie A, Shen B, Li S and Rao Z. Crystallization and preliminary X-ray diffraction analysis of FKBP12 complexed with a new neurotrophic ligand. Progress in Natural Science. 2003, 13:765-767 8. Li P, Shu C, Wu B, Ding Y, Shen B and Rao Z. Crystallization and preliminary X-ray diffraction analysis of FKBP52 N-terminal domain. Acta Cryst. D. 2002, 58:2168-2169 9. Li P, Wang L, Ding Y, Wu B, Shu C, Nie A, Li S, Shen B and Rao Z. Crystallization and preliminary X-ray diffraction analysis of FKBP12 complexed with a novel neurotrophic ligand, Protein and Peptide Letters. 2002, 5:459-463 9. Sun Y, Li X, Wu B, Sun P and Rao Z. Crystallization and Preliminary Crystallographic Analysis of Human eukaryotic translation initiation factor 5A (eIF-5A). Protein and Peptide Letters. 2005, 12:713-715 10. Zhao Q, Oin L, Jiang F, Wu B, Yue W, Xu F, Rong Z, Yuan H, Xie X, Gao Y, Bai C, Bartlam M, Pei X & Rao Z. Journal of Biological Chemistry. 2007, 282(1): 647-656 EXPERIMENTAL SKILLS • Molecular Biology 1. PCR techniques. 2. DNA recombination and expression of cloned genes in E. coli. 3. Site-directed mutagenesis and sub-cloning. • Protein Biochemistry 1. Purification and protein characterization of membrane proteins, especially G-protein coupled receptors (GPCRs). 2. CPM assays and analytical size-exclusion chromatography (aSEC) assays to analyze protein stability of GPCRs. 3. Expression, purification and characterization of recombinant proteins, including production of selenomethionine-substituted proteins. 4. Chromatographic techniques: ion-exchange chromatography, gel-filtration chromatography, hydrophobic interaction chromatography and affinity chromatography et al. 5. Purification with FPLC, AKTA explorer and AKTA purifier. 6. Characterization of proteins, nucleic acids, and small molecules using electrophoretic (native, non-reduced, reduced gel and Western blotting). • Protein Crystallization and Data Collection 1. Lipidic cubic phase (LCP) technique for membrane protein crystallization. 2. Fluorescence recovery after photobleaching (FRAP) assays to analyze membrane protein behavior in lipid cubic phase. 3. Vapor diffusion method using hanging drop or sitting drop. 4. Seeding techniques. 5. X-ray diffraction experiment and data collection. 6. Experienced in heavy-atom soaking. • Structure Determination 1. Phasing by molecular replacement (MR), multiple isomorphous replacement (MIR), single-wavelength anomalous dispersion (SAD) and multiple-wavelength anomalous dispersion (MAD) method. 2. Structure refinement and model building. 3. Structure presentation. 4. Much experience on crystallographic software: HKL2000, CNS, CCP4, COOT, Solve & Resolve, Molscript & Bobscript et al. RESEARCH EXPERIENCE My research experience covers bioinformatics analysis of the target proteins, gene cloning, protein expression in bacteria, membrane protein and soluble protein characterization, purification, and crystallization, structure determination and functional study of GPCRs and some human disease-related proteins. The two main projects are as follows: • Crystal structure of chemokine receptor CXCR4 (Research associate period) Chemokine receptors are critical regulators of cell migration in the context of immune surveillance, inflammation and development. The G protein-coupled chemokine receptor, CXCR4, is specifically implicated in cancer metastasis and HIV-1 infection. I solved five independent crystal structures of CXCR4 bound to an antagonist small molecule IT1t and a cyclic peptide CVX15 at 2.5-3.2 Å resolution. As the first structure of a protein-activated GPCR, CXCR4 structure shows many structural differences compared with the known GPCR structures. Except several differences in the 7TM helix bundle, all the CXCR4 structures lack the short α-helix VIII, which was thought to be a regular element of all Class A GPCRs. CXCR4 ligand binding pocket is more open, larger and located closer to the extracellular surface, which may correlate with the fact that CXCR4 natural ligand is CXCL12, a 10kDa protein. In all five CXCR4 structures, a similar parallel symmetric CXCR4 homodimer with an interface involving helices V and VI is observed. The dimer association is mostly driven by hydrophobic contacts, which persist throughout all five crystal forms, suggesting that these contacts present a biologically relevant homodimer interface. Many ligand-recetptor contacts in the co-crystal structures were reported to be important for CXCL12 binding and HIV-1 infectivity. The highly positively charged peptide ligand CVX15 and the small molecule ligand IT1t may trace the path of the N-terminal signaling peptide of CXCL12 and the V3 loop of HIV-1 gp120, which were reported to penetrate the CXCR4 binding pocket. The CXCR4 co-crystal structures provide new clues about the interactions between CXCR4 and its natural ligand CXCR12 and with the HIV-1 glycoprotein gp120. The paper about this work was accepted by Science as a research article. • Crystal structure of human FKBP52 (Ph. D period) Immunophilins are proteins possessing peptidylprolyl isomerase (PPIase) domains that bind immunosuppressant drugs. FKBP52, which can bind FK506 and possess PPIase activity, is an important immunophilin involved in the heterocomplex of steroid receptors with Hsp90. I cloned, purified and crystallized two overlapped fragments [N(1-260) and C(145-459)] of FKBP52 and solved the structures, as well as the complex of C(145-459) and a C-terminal pentapeptide MEEVD from Hsp90. Based on the structure of these two overlapped fragments, the complete putative structure of FKBP52 was defined. The structure of FKBP52 is architecturally similar to another immunophilin FKBP51 except that the orientations between different domains are quite different. A detailed structural analysis of FKBP52 and the complex with the MEEVD peptide has revealed the essential basis for the loss of PPIase activity of the FK2 domain and the key residues for Hsp90 binding. Key structural differences between FKBP52 and FKBP51, including the relative orientations of the four domains and some important residue substitutions, account for the differential functions of FKBPs. Based on the structures of FKBPs and ligand binding domain (LBD) of GR, I clarified several features of interaction between components of the GR complexes and proposed a model of GR·Hsp90·FKBPs heterocomplex assembly. In this model, the FKBPs complexes have different stability and preference for hormone and FKBP51 is a negative feedback factor in the pathway of FKBP52-mediated hormone signaling. This research work has been published in PNAS (2004, 101:8348-8353). • Other projects I involved in: Crystallographic studies of influenza virus polymerase, pyrohposphatase RS21-C6, HLA-G, tumor necrosis factor hBLyS, calcineurin B