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Boris Brumshtein

Joined 28 October 2007
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[[Image:BorisBrumshtein-face-picture.png|thumb|right|190px| Year:2007]]
==CV==
==CV==


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===Education===
===Education===


2003 - present  Direct PhD program at the Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.  
*''2003 - present'' Direct PhD program at the Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.  
Estimated graduation - summer 2008. Scientific advisors: Prof. Joel L. Sussman, Prof. Israel Silman, Prof. Anthony H. Futerman  
Estimated graduation - summer 2008. Scientific advisors: Prof. Joel L. Sussman, Prof. Israel Silman, Prof. Anthony H. Futerman  
Thesis topic: "Structural and biochemical studies on acid-beta-glucosidase, the defected protein in Gaucher disease".
Thesis topic: "Structural and biochemical studies on acid-beta-glucosidase, the defective protein in Gaucher disease".
 
*''2002-2003'' MSc student, Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.
2002-2003 MSc student, Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.
Scientific advisor: Prof. Joel Sussman.
Scientific advisor: Prof. Joel Sussman.
 
*''1999-2002'' BA in Biochemistry, Technion, Israel Institute of Science and Technology, Faculty of Chemistry, cum laude.
1999-2002 BA in Biochemistry, Technion, Israel Institute of Science and Technology, Faculty of Chemistry, cum laude.  


===Qualifications===
===Qualifications===
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====Crystallography====
====Crystallography====
   
   
- Protein crystallization utilizing microbatch crystallization, IMPAX I-5, Oryx and Mosquito crystallization robots.   
*Protein crystallization utilizing microbatch crystallization, IMPAX I-5, Oryx and Mosquito crystallization robots.   
*''Ab initio'' crystallization and crystals optimization. Various crystallization techniques, such as: soaking, co-crystallization, crystallization in presence of heavy metals for phasing, etc.
*Crystal handling and manipulations, micro-manipulations with ZEISS microscope/manipulator system.
*Data acquisition using Rigaku Raxis 4/4++ or synchrotron x-ray sources. 
*X-ray data processing software: XDS, HKL2000. 
*Protein structure determination using MR, SIR, MIR, MAD, SIRAS approaches with CNS, SHELX and CCP4 software suits. 
*Molecular modeling, visualization and analysis software: PyMol, Coot, XtalView, O, CCP4, APBS. 
*Docking and small molecule/drug database mining with AutoDock - drug docking and virtual screening, Cambridge Structure Database (CSB), ZINC, ChemOffice.


- ''Ab initio'' crystallization and crystals optimization. Various crystallization techniques, such as: soaking, co-crystallization, crystallization in presence of heavy metals for phasing, etc.
====Molecular biology and biochemistry====
*Protein cloning and expression in E.Coli. 
*Biochemical and protein purification techniques: HPLC, FPLC, Ni-columns, Ion exchange columns, acryl-amide gels, size exclusion, affinity chromatography, gradient and analytical ultracentrifugation, ultra-filtration, Mini Prep cell, etc.
*Drug-Protein interaction analysis employing various structural, biochemical and biophysical assays such as: enzymatic assays, ELIZA, circular dichroism, dynamic light scattering, mass-spec, BiaCore, analytical ultracentrifugation, etc.


- Crystal handling and manipulations, micro-manipulations with ZEISS microscope/manipulator system.
====Programming languages====


- Data acquisition using Rigaku Raxis 4/4++ or synchrotron x-ray sources.
*Unix, C, Perl, Matlab, Java (basic programming).


- X-ray data processing software: XDS, HKL2000. 


- Protein structure determination using MR, SIR, MIR, MAD, SIRAS approaches with CNS, SHELX and CCP4 software suits.   
==LIST OF PUBLICATIONS==
#Kacher Y, Brumshtein B, Boldin-Adamsky S, Toker L, Shainskaya A, Silman I, Sussman JL, Futerman AH. Acid-beta-glucosidase: insights from structural analysis and relevance to Gaucher disease therapy. Biol Chem. 2008 Sep 11. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/18783340 18783340]
#Harel M, Brumshtein B, Meged R, Dvir H, Ravelli RB, McCarthy A, Toker L, Silman I, Sussman JL. 3-D structure of serum paraoxonase 1 sheds light on its activity, stability, solubility and crystallizability. Arh Hig Rada Toksikol. 2007 Sep;58(3):347-53. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17913690 17913690]
# Brumshtein B, Greenblatt HM, Butters TD, Shaaltiel Y, Aviezer D, Silman I, Futerman AH, Sussman JL. Crystal structures of complexes of N-butyl- and N-nonyl-deoxynojirimycin bound to acid-beta -glucosidase: Insights into the mechanism of chemical chaperone action in Gaucher disease. J.Biol. Chem., Vol. 282, Issue 39, 29052-29058, (2007). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17666401 17666401]
# Brumshtein, B., Wormald, M. R., Silman, I., Futerman, A. H. & Sussman, J. L. Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. Acta Crystallogr D Biol Crystallogr 62, 1458-65 (2006). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17139081 17139081]
# Rydberg EH, Brumshtein B, Greenblatt HM, Wong DM, Shaya D, Williams LD, Carlier PR, Pang YP, Silman I, Sussman JL. Complexes of alkylene-linked tacrine dimers with Torpedo californica acetylcholinesterase: Binding of Bis5- tacrine produces a dramatic rearrangement in the active-site gorge. J Med Chem 49, 5491-500 (2006). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16942022 16942022]
# Harel M, Hyatt JL, Brumshtein B, Morton CL, Wadkins RM, Silman I, Sussman JL, Potter PM. The 3D structure of the anticancer prodrug CPT-11 with Torpedo californica acetylcholinesterase rationalizes its inhibitory action on AChE and its hydrolysis by butyrylcholinesterase and carboxylesterase. Chem Biol Interact 157-158, 153-7 (2005). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16289500 16289500]
# Hyatt JL, Tsurkan L, Morton CL, Yoon KJ, Harel M, Brumshtein B, Silman I, Sussman JL, Wadkins RM, Potter PM. Inhibition of acetylcholinesterase by the anticancer prodrug CPT-11. Chem Biol Interact 157-158, 247-52 (2005). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16257398 16257398]
# Harel M, Hyatt JL, Brumshtein B, Morton CL, Yoon KJ, Wadkins RM, Silman I, Sussman JL, Potter PM. The crystal structure of the complex of the anticancer prodrug 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin (CPT- 11) with Torpedo californica acetylcholinesterase provides a molecular explanation for its cholinergic action. Mol Pharmacol 67, 1874-81 (2005).  PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15772291 15772291]
# Harel M, Aharoni A, Gaidukov L, Brumshtein B, Khersonsky O, Meged R, Dvir H, Ravelli RB, McCarthy A, Toker L, Silman I, Sussman JL, Tawfik DS. Structure and evolution of the serum paraoxonase family of detoxifying and anti-atherosclerotic enzymes. Nat Struct Mol Biol 11, 412-9 (2004). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15098021 15098021]
# Adir, N., Rukhman, V., Brumshtein, B. & Anati, R. Preliminary X-ray crystallographic analysis of a soluble form of MntC, a periplasmic manganese- binding component of an ABC-type Mn transporter from Synechocystis sp. PCC 6803. Acta Crystallogr D Biol Crystallogr 58, 1476-8 (2002). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/12198307 12198307]
   


- Molecular modeling, visualization and analysis software: PyMol, Coot, XtalView, O, CCP4, APBS.
==PROTEIN DATABANK DEPOSITIONS==
 
- Docking and small molecule/drug database mining with AutoDock - drug docking and virtual screening, Cambridge Structure Database (CSB), ZINC, ChemOffice.
#[[1odc]] STRUCTURE OF ACETYLCHOLINESTERASE (E.C. 3.1.1.7) COMPLEXED WITH N-4'-QUINOLYL-N'-9"-(1",2",3",4"- TETRAHYDROACRIDINYL)-1,8-DIAMINOOCTANE AT 2.2A RESOLUTION
#[[1u65]] Ache W. CPT-11
#[[1ut6]] STRUCTURE OF ACETYLCHOLINESTERASE (E.C. 3.1.1.7) COMPLEXED WITH N-9-(1',2',3',4'-TETRAHYDROACRIDINYL)-1,8-DIAMINOOCTANE AT 2.4 ANGSTROMS RESOLUTION
#[[1v04]] SERUM PARAOXONASE BY DIRECTED EVOLUTION
#[[2ckm]] TORPEDO CALIFORNICA ACETYLCHOLINESTERASE COMPLEXED WITH ALKYLENE-LINKED BIS-TACRINE DIMER (7 CARBON LINKER)
#[[2cmf]] TORPEDO CALIFORNICA ACETYLCHOLINESTERASE COMPLEXED WITH ALKYLENE-LINKED BIS-TACRINE DIMER (5 CARBON LINKER)  
#[[2j25]] PARTIALLY DEGLYCOSYLATED GLUCOCERAMIDASE
#[[2v3d]] ACID-BETA-GLUCOSIDASE WITH N-BUTYL-DEOXYNOJIRIMYCIN
#[[2v3e]] ACID-BETA-GLUCOSIDASE WITH N-NONYL-DEOXYNOJIRIMYCIN


====Molecular biology and biochemistry====


- Protein cloning and expression in E.Coli. 
==MY CURRENT RESEARCH==


- Biochemical and protein purification techniques: HPLC, FPLC, Ni-columns, Ion
My PhD research thesis focuses on the enzyme acid-beta-glucosidase, with the
exchange columns, acryl-amide gels, size exclusion, affinity chromatography,
emphasis on the enzyme's structure and its interactions with various ligands and
gradient and analytical ultracentrifugation, ultra-filtration, Mini Prep cell, etc.
their pharmacological potential.  


- Drug-Protein interaction analysis employing various structural, biochemical and
The enzyme is involved in a Gaucher disease, which is the most common
biophysical assays such as: enzymatic assays, ELIZA, circular dichroism, dynamic
lysosomal storage disease, and is associated with mutations in the gene
light scattering, mass-spec, BiaCore, analytical ultracentrifugation, etc.
coding for the enzyme acid-beta-glucosidase.  


====Programming languages====
There are ~200 known mis-sense mutations, which result in substitution of amino acids in the protein.
Some mutations cause complete deactivation of the enzyme; others impair its stability, and some
affect both activity and stability.


-Unix, C, Perl, Matlab, Java (basic programming).
There are three known phenotypes of the disease: a mild, severe and acute.
The acute phenotype is neuropathic, while severe and mild symptoms are caused by accumulation of
glucoceramide in macrophage cells resulting in bone atrophy, spleen enlargement, etc.  


We have solved structures of recombinant acid-beta-glucosidase expressed in both mammalian
and plant cell cultures, as well as of complexes with molecules, which have been shown to inhibit
glucoceramide synthase and in a less extent acid-beta-glucosidase. Yet these molecules were shown
to stabilize several mutant forms of the mutant enzyme in Gaucher disease.


==LIST OF PUBLICATIONS==
We have shown that the molecular basis for the stabilization of acid-beta-glucosidase by these small molecules
is by binding to the active site and contributing additional non-covalent bonds that stabilize the enzyme's overall structure.


# Brumshtein B, Greenblatt HM, Butters TD, Shaaltiel Y, Aviezer D, Silman I, Futerman AH, Sussman JL. Crystal structures of complexes of N-butyl- and N-nonyl-deoxynojirimycin bound to acid-beta -glucosidase: Insights into the mechanism of chemical chaperone action in Gaucher disease. J.Biol. Chem., Vol. 282, Issue 39, 29052-29058, (2007).
# Brumshtein, B., Wormald, M. R., Silman, I., Futerman, A. H. & Sussman, J. L. Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. Acta Crystallogr D Biol Crystallogr 62, 1458-65 (2006).
# Rydberg EH, Brumshtein B, Greenblatt HM, Wong DM, Shaya D, Williams LD, Carlier PR, Pang YP, Silman I, Sussman JL. Complexes of alkylene-linked tacrine dimers with Torpedo californica acetylcholinesterase: Binding of Bis5- tacrine produces a dramatic rearrangement in the active-site gorge. J Med Chem 49, 5491-500 (2006).
# Harel M, Hyatt JL, Brumshtein B, Morton CL, Wadkins RM, Silman I, Sussman JL, Potter PM. The 3D structure of the anticancer prodrug CPT-11 with Torpedo californica acetylcholinesterase rationalizes its inhibitory action on AChE and its hydrolysis by butyrylcholinesterase and carboxylesterase. Chem Biol Interact 157-158, 153-7 (2005).
# Hyatt JL, Tsurkan L, Morton CL, Yoon KJ, Harel M, Brumshtein B, Silman I, Sussman JL, Wadkins RM, Potter PM. Inhibition of acetylcholinesterase by the anticancer prodrug CPT-11. Chem Biol Interact 157-158, 247-52 (2005).
# Harel M, Hyatt JL, Brumshtein B, Morton CL, Yoon KJ, Wadkins RM, Silman I, Sussman JL, Potter PM. The crystal structure of the complex of the anticancer prodrug 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin (CPT- 11) with Torpedo californica acetylcholinesterase provides a molecular explanation for its cholinergic action. Mol Pharmacol 67, 1874-81 (2005). 
# Harel M, Aharoni A, Gaidukov L, Brumshtein B, Khersonsky O, Meged R, Dvir H, Ravelli RB, McCarthy A, Toker L, Silman I, Sussman JL, Tawfik DS. Structure and evolution of the serum paraoxonase family of detoxifying and anti-atherosclerotic enzymes. Nat Struct Mol Biol 11, 412-9 (2004).
# Adir, N., Rukhman, V., Brumshtein, B. & Anati, R. Preliminary X-ray crystallographic analysis of a soluble form of MntC, a periplasmic manganese- binding component of an ABC-type Mn transporter from Synechocystis sp. PCC 6803. Acta Crystallogr D Biol Crystallogr 58, 1476-8 (2002). PMID:[http://www.ncbi.nlm.nih.gov/pubmed/12198307 12198307]


The research is been performed in laboratories of [http://www.weizmann.ac.il/sb/faculty_pages/Sussman/ Prof. Joel L. Sussman], [http://www.weizmann.ac.il/neurobiology/faculty.html Prof. Israel Silman] and [http://www.weizmann.ac.il/Biological_Chemistry/scientist/futerman/ Prof. Tony H. Futerman],
[http://www.weizmann.ac.il/ Weizmann Institute of Science]


The page is always under construction ; last update [[User:Boris Brumshtein|Boris Brumshtein]] 14:12, 8 June 2008 (IDT)


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