Masoud Vedadi, Ph.D.

Principal Investigator, Molecular Biophysics

Tel: 416 946 0897
e-mail: mvedadi@uhnres.utoronto.ca

Masoud received his Ph.D. from McGill University (1997) and worked as a postdoctoral fellow in the Department of Biochemistry in Concordia University, Canada, focusing on structural and functional studies of enzymes (1997-1999).

Masoud then joined Affinium Pharmaceuticals where he worked as Associate Director of Protein Science. There, he and his team were involved in purifying, characterizing and crystallizing hundreds of proteins as well as developing novel methods of protein purification and characterization (1999-2003). He later joined the Structural Genomics Consortium in 2003 as head of the Biophysics group.

Masoud and his team characterize proteins produced by the SGC scientists in Toronto and also focus on developing new high throughput biophysical and biochemical characterization techniques. By screening proteins against customized chemical libraries, they identify ligands that promote crystallization and improve crystal quality. They also screen for buffer conditions that reduce protein aggregation and enable them to further concentrate problematic proteins. Creating and comparing chemical profiles for members of each family of protein along with functional studies, they investigate the structure-function relationship and identify unique inhibitors that can discriminate among members of each family of protein. Identifying human SNPs that affect human health by employing high throughput screening techniques is also an ongoing project in his group.

Team Members (left to right)

• Masoud Vedadi,
• Guillermo Senisterra, Ph.D., Research Associate
• Abdellah Allali-Hassani, Ph.D., Sr. Research Associate
• Gregory Wasney, M.Sc., Laboratory Technician
• Taraneh Hajian, M.Sc., Laboratory Technician
• Irene Chau, M.Sc., Laboratory Technician
• Alena Siarheyeva, Ph.D., Research Associate
  
  
  

Small Molecule Screening Workshop

SGC Toronto July 8-9^th, 2010;

As part of the SGC’s ongoing program in structural genomics of human and malaria proteins we are also identifying small molecules that bind and stabilize our proteins, thereby promoting crystallization. We have implemented two screening platforms, based on either fluorimetry or static light scattering, to measure the increase in protein thermal stability upon binding of a ligand without the need to monitor enzyme activity. Our small molecule libraries comprise many bioactive compounds such as nucleotides, cofactors, substrates and inhibitors, as well as FDA-approved drugs and drug-like compounds. Identification of small molecules that bind to proteins can provide valuable information on protein specificity, activity, as well as a practical means to facilitate 3D structure determination. This workshop aims to provide an opportunity for interested scientists to learn how to implement these techniques, free of charge, and enable them to screen a protein of interest against selected compounds during the workshop. This is a two day workshop and will be held at the SGC labs, University of Toronto. Please contact us by e-mail if you need more information.

Contact: mvedadi@uhnres.utoronto.ca

Publications

Guillermo A. Senisterra, Hamed Ghanei, Galina Khutoreskaya, Elena Dobrovetsky, Aled M. Edwards, Gilbert G. Privé and Masoud Vedadi (2010) Assessing the stability of membrane proteins to detect ligand binding using differential static light scattering. J. Biomol. Screen. In Press.

Amy Quinn, Abdellah Allali-Hassani, Masoud Vedadi, Anton Simeonov (2010) A Chemiluminescence-based Method for Identification of Histone Lysine Methyltransferase Inhibitors. Molecular BioSystems. In press.

Wu H, Min J, Lunin VV, Antoshenko T, Dombrovski L, Zeng H, Allali-Hassani A, Campagna-Slater V, Vedadi M, Arrowsmith CH, Plotnikov AN, Schapira M. (2010) Structural biology of human H3K9 methyltransferases. PLoS One. 11;5(1):e8570.

Tim J. Wigle, J. Martin Herold, Guillermo Senisterra, Masoud Vedadi, Dmitri B. Kireev, Cheyl H. Arrowsmith, Stephen V. Frye, and William P. Janzen. (2010) Screening for Inhibitors of Low-Affinity Epigenetic Peptide-Protein Interactions: An AlphaScreen™-Based Assay for Antagonists of Methyl-Lysine Binding Proteins. J. Biomol. Screen. 15(1):62-71.

Feng Liu, Xin Chen, Abdellah Allali-Hassani, Amy Quinn, Gregory Wasney, Aiping Dong, Dalia Barsyte, Ivona Kozieradzki, Guillermo Senisterra, Irene Chau, Alena Siarheyeva, Dmitri Kireev, Ajit Jadhav, Martin Herold, Stephen Frye, Cheryl Arrowsmith, Peter Brown, Anton Simeonov, Masoud Vedadi, Jian Jin. (2009) Discovery of a 2,4-Diamino-7-aminoalkoxy-quinazoline as a Potent and Selective Inhibitor of Histone Lysine Methyltransferase G9a. J. Med. Chem. 52(24):7950-3.

Abdellah Allali-Hassani, Gregory A. Wasney, Irene Chau, Bum Soo Hong, Guillermo Senisterra, Peter Loppnau, Zhen Shi, John Moult, Aled M. Edwards, Cheryl H. Arrowsmith, Hee Won Park, Matthieu Schapira and Masoud Vedadi (2009) A survey of proteins encoded by non-synonymous SNPs reveals a significant fraction with altered stability and activity. Biochem. J. 424(1):15-26

Eryilmaz J, Pan P, Amaya MF, Allali-Hassani A, Dong A, Adams-Cioaba MA, Mackenzie F, Vedadi M, Min J. (2009) Structural studies of a four-MBT repeat protein MBTD1. PLoS One. Oct 20;4(10):e7274.

Min T, Vedadi M, Watson D, Wasney G, Munger C, Cygler M, Matte A, Young N. (2009) Specificity of the Campylobacter jejuni Adhesin PEB3 for Phosphates and Structural Differences among its Ligand Complexes. Biochemistry. 48(14):3057-67

Guo Y, Nady N, Qi C, Allali-Hassani A, Zhu H, Pan P, Adams-Cioaba MA, Amaya MF, Dong A, Vedadi M, Schapira M, Read RJ, Arrowsmith CH, Min J. (2009) Methylation-state-specific recognition of histones by the MBT repeat protein L3MBTL2. Nucleic Acids Res. 37(7):2204-10.

Huang X, Finerty P Jr, Walker JR, Butler-Cole C, Vedadi M, Schapira M, Parker SA, Turk BE, Thompson DA, Dhe-Paganon S. (2008) Structural insights into the inhibited states of the Mer receptor tyrosine kinase. J Struct Biol. 165:88-96.

Guillermo A. Senisterra, Bum Soo Hong, Hee-Won Park and Masoud Vedadi (2008) Application of high throughput isothermal denaturation to assess protein stability and screen for ligands. J Biomol Screen. 13(5):337-42.

Min J, Allali-Hassani A, Nady N, Qi C, Ouyang H, Liu Y, MacKenzie F, Vedadi M, Arrowsmith CH (2008) L3MBTL1 recognition of mono- and dimethylated histones. Nat Struct Mol Biol.14(12):1229-30.

Schuetz A, Min J, Allali-Hassani A, Schapira M, Shuen M, Loppnau P, Mazitschek R, Kwiatkowski NP, Lewis TA, Maglathin RL, McLean TH, Bochkarev A, Plotnikov AN, Vedadi M, Arrowsmith CH. (2008) Human HDAC7 harbors a class IIa histone deacetylase-specific zinc binding motif and cryptic deacetylase activity. J Biol Chem. 283(17):11355-63.

Niesen F, Berglund H, and Vedadi M. (2007) The use of Differential Scanning Fluorimetry to detect ligand interactions that promote protein stability. Nature Protocols. 2(9):2212-21.

Hong BS, Senisterra G, Rabeh WM, Vedadi M, Leonardi R, Zhang YM, Rock CO, Jackowski S, Park HW (2007) Crystal structures of human pantothenate kinases. Insights into allosteric regulation and mutations linked to a neurodegeneration disorder. J Biol Chem. 282(38):27984-93.

Allali-Hassani A, Pan PW, Dombrovski L, Najmanovich R, Tempel W, Dong A, Loppnau P, Martin F, Thornton J, Edwards AM, Bochkarev A, Plotnikov AN, Vedadi M, Arrowsmith CH. (2007) Structural and Chemical Profiling of the Human Cytosolic Sulfotransferases. PLoS Biology. 5(5): e97.

Vedadi M, Lew J, Artz J, Amani M, Zhao Y, Dong A, Wasney GA, Gao M, Hills T, Brokx S, Qiu W, Sharma S, Diassiti A, Alam Z, Melone M, Mulichak A, Wernimont A, Bray J, Loppnau P, Plotnikova O, Newberry K, Sundararajan E, Houston S, Walker J, Tempel W, Bochkarev A, Kozieradzki I, Edwards A, Arrowsmith C, Roos D, Kain K, Hui R. (2007) Genome-scale protein expression and structural biology of Plasmodium falciparum and related Apicomplexan organisms. Mol Biochem Parasitol. 151(1):100-10.

Schuetz A, Min J, Antoshenko T, Wang CL, Allali-Hassani A, Dong A,Loppnau P, Vedadi M, Bochkarev A, Sternglanz R, Plotnikov AN. (2007) Structural Basis of Inhibition of the Human NAD(+)-Dependent Deacetylase SIRT5 by Suramin. Structure. 15(3):377-89.

Wu H, Horton JR, Battaile K, Allali-Hassani A, Martin F, Zeng H, Loppnau P, Vedadi M, Bochkarev A, Plotnikov AN, Cheng X. (2007) Structural basis of allele variation of human thiopurine-S-methyltransferase. Proteins. 67(1):198-208.

Lorca GL, Ezersky A, Lunin VV, Walker JR, Altamentova S, Evdokimova E, Vedadi M, Bochkarev A, Savchenko A.(2007) Glyoxylate and pyruvate are antagonistic effectors of the Escherichia coli IclR transcriptional regulator. J Biol Chem. 282:16476-91.

Dong A, Xu X, Edwards AM, Chang C, Chruszcz M, Cuff M, Cymborowski M, Di Leo R, Egorova O, Evdokimova E, Filippova E, Gu J, Guthrie J, Ignatchenko A, Joachimiak A, Klostermann N, Kim Y, Korniyenko Y, Minor W, Que Q, Savchenko A, Skarina T, Tan K, Yakunin A, Yee A, Yim V, Zhang R, Zheng H, Akutsu M, Arrowsmith C, Avvakumov GV, Bochkarev A, Dahlgren LG, Dhe-Paganon S, Dimov S, Dombrovski L, Finerty P Jr, Flodin S, Flores A, Gräslund S, Hammerström M, Herman MD, Hong BS, Hui R, Johansson I, Liu Y, Nilsson M, Nedyalkova L, Nordlund P, Nyman T, Min J, Ouyang H, Park HW, Qi C, Rabeh W, Shen L, Shen Y, Sukumard D, Tempel W, Tong Y, Tresagues L, Vedadi M, Walker JR, Weigelt J, Welin M, Wu H, Xiao T, Zeng H, Zhu H. (2007) In situ proteolysis for protein crystallization and structure determination. Nat Methods. 4:1019-21.

Vedadi M, Niesen FH, Allali-Hassani A, Fedorov OY, Finerty PJ Jr, Wasney GA, Yeung R, Arrowsmith C, Ball LJ, Berglund H, Hui R, Marsden BD, Nordlund P, Sundstrom M, Weigelt J, Edwards AM. (2006) Chemical screening methods to identify ligands that promote protein stability, protein crystallization, and structure determination. Proc Natl Acad Sci U S A. 103 (43):15835-40.

Schuetz A, Allali-Hassani A, Martin F, Loppnau P, Vedadi M, Bochkarev A, Plotnikov AN, Arrowsmith CH, Min J. (2006) Structural basis for molecular recognition and presentation of histone H3 by WDR5. EMBO J. 25(18):4245-52.

Senisterra GA, Markin E, Yamazaki K, Hui R, Vedadi M and Awrey DE (2006) Screening for ligands using a generic and high-throughput light-scattering-based assay. J Biomol Screen; 11(8): 940-8

Kimber MS, Martin F, Lu Y, Houston S, Vedadi M, Dharamsi A, Fiebig KM, Schmid M, Rock CO (2004) The structure of (3R)-hydroxyacyl-acyl carrier protein dehydratase (FabZ) from Pseudomonas aeruginosa. J Biol Chem. 279:52593-602

Kimber, M, Vallee, F, Houston, S, Necakov, A, Skarina, T, Evdokimova, E, Beasley, S, Christendat, D, Savchenko, A, Arrowsmith, C, Vedadi, M, Gerstein, M, and Edwards, A. (2003) Data mining crystallization databases: Knowledge-based approaches to optimise protein crystal screens. PROTEINS: Structure, Function and Genetics. 51:562-568.

Khamessan,A, Naghibalhossaini F, Vedadi,M,and Johnstone,R.M. (2001) Mutated gama-actin restores growth to a yeast amino acid transport defective mutant. J Cell Physiol.186: 124-35.

Edwards AM, Arrowsmith CH, Christendat D, Dharamsi A, Friesen JD, Greenblatt JF, Vedadi M. (2000) Protein production: feeding the crystallographers and NMR spectroscopists. Nat Struct Biol. l:970-2

Ahvazi B, Coulombe R, Delarge M, Vedadi M,Zhang L, Meighen E, and Vrielink, A. (2000) Crystal structure of the NADP+ dependant aldehyde dehydrogenase from Vibrio harveyi. : structural implications for cofactor specificity and affinity. Biochem J. 349:853-61.

Vedadi M, Barriault D, Sylvestre, M and Powlowski, J (2000) Active site residues of cis-2,3-dihydro-2,3-dihydroxybiphenyl dehydrogenase from Comamonas testosteroni strain B-356. Biochemistry. 39:5028-34.

Barriault D, Vedadi M, Powlowski J,and Sylvestre M (1999) Cis-2,3-dihydro-2,3-dihydroxybiphenyl dehydrogenase and cis-1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase catalyze dehydrogenation of the same range of substrates. Biochem. Biophys. Res. Commun. 260:181-7.

Vedadi M, Vrielink A, Meighen E (1997) Involvement of conserved glycine residues, 229 and 234, of V. harveyi aldehyde dehydrogenase in activity and nucleotide binding. Biochem. Biophys. Res. Commun. 238(2), 448-451.

Vedadi M, Croteau N, Delarge M, Vrielink A, Meighen E (1997) Structural and functional studies of an NADP+-specific aldehyde dehydrogenase from the luminescent marine bacterium V. harveyi. Adv.Exp.Med.Biol. 414,269-275.

Vedadi M, Meighen E (1997) Critical glutamic acid residues affecting the mechanism and nucleotide specificity of V. harveyi aldehyde dehydrogenase. Eur.J.Biochem.246, 698-704.

Vedadi M and Meighen. (1995) Is fatty aldehyde dehydrogenase expression under similar control as the bioluminescent system in V. harveyi in Bioluminescence and Chemiluminescence.[Campbell,A, Kricka,LJ, Stanley,PE. (Ed)] pp 556-559, John Wiley & Sons, New York.

Vedadi M, Szittner R, Smillie L, and Meighen E(1995) Involvement of cysteine 289 in the catalytic activity of an NADP+-specific aldehyde dehydrogenase from V. harveyi. Biochemistry. 34,16725-16732.

Some of standard SOP's employed by the biophysics group:

• Compound storage
• Screening by FluoDia T70
• Screening by Stratagene Mx3005P