1) “Develop high-throughput assay for screening specific inhibitors of ZAP-70.” ZAP-70 is a non-receptor tyrosine kinase that normally expresses in T cells and natural killer cells and has a critical role in the initiation of T-cell signaling. As tyrosine kinases are usually very important targets for cancer chemotherapy, I am setting out to develop high-throughput assays for the purpose of screening specific inhibitors of ZAP-70. I am also trying to use the bacterial expression system to produce ZAP-70 protein, as it is much cheaper and more efficient than the baculovirus expression system.
2) “Investigate the regulation mechanism of Ack1 activity.” Ack1 is a non-receptor tyrosine that activates downstream of melanoma chondroitin sulfate proteoglycan, thereby regulating tumor cell motility and invasion. The domain organization of Ack1 is very different from the other non-receptor tyrosine kinases, which consists of an N-terminal SAM domain, a kinase domain, a SH3 domain, a CDC42 binding domain, and a C-terminal domain homologous to the epidermal growth factor receptor-binding domain of Gene33 (also called MIG6). Despite the important role in cell signaling, the activation and regulation mechanism of Ack1 is not well understood. I am exploring how X-ray crystallography methods can be used in combination with other biochemical and biophysical techniques to illustrate the regulation mechanism of Ack1 activity.
Impact in China
The pharmaceutical industry has experienced dramatic and explosive growth in recent years, and there is increasing interest in China’s potential to supply novel preclinical and clinical compounds. With the knowledge and techniques learned in Berkeley, I hope that I can contribute to drug discovery research in China.
Yang JP, Ma XX, He YX, Li WF, Kang Y, Bao R, Chen Y, Zhou CZ. Crystal structure of the 30 K protein from the silkworm Bombyx mori reveals a new member of the β-trefoil superfamily. J Struct Biol. 2011 Jul;175(1):97-103.
Du Y, Shi WW, He YX, Yang YH, Zhou CZ, Chen Y. Structures of the substrate-binding protein provide insights into the multiple compatible solute binding specificities of the Bacillus subtilis ABC transporter OpuC. Biochem J. 2011 Jun 1;436(2):283-9.
Du Y, He YX, Zhang ZY, Yang YH, Shi WW, Frolet C, Di Guilmi AM, Vernet T, Zhou CZ, Chen Y. Crystal structure of the mucin-binding domain of Spr1345 from Streptococcus pneumoniae. J Struct Biol. 2011 Apr;174(1):252-7.
Zhang X, Teng YB, Liu JP, He YX, Zhou K, Chen Y, Zhou CZ. Structural insights into the catalytic mechanism of the yeast pyridoxal 5-phosphate synthase Snz1. Biochem J. 2010 Nov 25;432(3):445-50.
Zhao MX, Jiang YL, He YX, Chen YF, Teng YB, Chen Y, Zhang CC, Zhou CZ. Structural basis for the allosteric control of the global transcription factor NtcA by the nitrogen starvation signal 2-oxoglutarate. Proc Natl Acad Sci U S A. 2010 Jul 13;107(28):12487-92.
Du Y, He YX, Gaowa S, Zhang X, Chen Y, Zhang SC, Zhou CZ. Crystal structures of the apo and GDP-bound forms of a cupin-like protein BbDUF985 from Branchiostoma belcheri tsingtauense. Proteins. 2010 Sep;78(12):2714-9.
Luo M, Jiang YL, Ma XX, Tang YJ, He YX, Yu J, Zhang RG, Chen Y, Zhou CZ. Structural and biochemical characterization of yeast monothiol glutaredoxin Grx6. J Mol Biol. 2010 May 14;398(4):614-22.
Zhang NN, He YX, Li WF, Teng YB, Yu J, Chen Y, Zhou CZ. Crystal structures of holo and Cu-deficient Cu/Zn-SOD from the silkworm Bombyx mori and the implications in amyotrophic lateral sclerosis. Proteins. 2010Jun;78(8):1999-2004.
He YX, Huang L, Xue Y, Fei X, Teng YB, Rubin-Pitel SB, Zhao H, Zhou CZ. Crystal structure and computational analyses provide insights into the catalytic mechanism of 2,4-diacetylphloroglucinol hydrolase PhlG from Pseudomonas fluorescens. J Biol Chem. 2010 Feb 12;285(7):4603-11.
Teng YB, Jiang YL, He YX, He WW, Lian FM, Chen Y, Zhou CZ. Structural insights into the substrate tunnel of Saccharomyces cerevisiae carbonic anhydrase Nce103. BMC Struct Biol. 2009 Oct 24;9:67.
Ma XX, Jiang YL, He YX, Bao R, Chen Y, Zhou CZ. Structures of yeast glutathione-S-transferase Gtt2 reveal a new catalytic type of GST family. EMBO Rep. 2009 Dec;10(12):1320-6.
Teng YB, Ma XX, He YX, Jiang YL, Du J, Xiang C, Chen Y, Zhou CZ. Crystal structure of Arabidopsis translation initiation factor eIF-5A2. Proteins. 2009 Nov 15;77(3):736-40.
Wang Y, He YX, Yu J, Zhou CZ. Cloning, overproduction, purification, crystallization and preliminary X-ray diffraction analysis of yeast glutaredoxin Grx5. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Jun 1;65(Pt 6):651-3.
He YX, Gui L, Liu YZ, Du Y, Zhou Y, Li P, Zhou CZ. Crystal structure of Saccharomyces cerevisiae glutamine synthetase Gln1 suggests a nanotube-like supramolecular assembly. Proteins. 2009 Jul;76(1):249-54.
Huang L, Sun F, Liang C, He YX, Bao R, Liu L, Zhou CZ. Crystal structure of LZ-8 from the medicinal fungus Ganoderma lucidium. Proteins. 2009 May 1;75(2):524-7.
Zhang WJ, He YX, Yang Z, Yu J, Chen Y, Zhou CZ. Crystal structure of glutathione-dependent phospholipid peroxidase Hyr1 from the yeast Saccharomyces cerevisiae. Proteins. 2008 Dec;73(4):1058-62.
Wang S, He YX, Bao R, Teng YB, Ye BP, Zhou CZ. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of hypothetical protein SCO4226 from Streptomyces coelicolor A3(2). Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Sep 1;64(Pt 9):847-50.