Three-dimensional structure determination of proteins related to human health in their functional context at The Israel Structural Proteomics Center (ISPC). This paper was presented at ICCBM10.
The principal goal of the Israel Structural Proteomics Center (ISPC) is to determine the structures of proteins related to human health in their functional context. Emphasis is on the solution of structures of proteins complexed with their natural partner proteins and/or with DNA. To date, the ISPC has solved the structures of 14 proteins, including two protein complexes. It has adopted automated high-throughput (HTP) cloning and expression techniques and is now expressing in Escherichia coli, Pichia pastoris and baculovirus, and in a cell-free E. coli system. Protein expression in E. coli is the primary system of choice in which different parameters are tested in parallel. Much effort is being devoted to development of automated refolding of proteins expressed as inclusion bodies in E. coli. The current procedure utilizes tagged proteins from which the tag can subsequently be removed by TEV protease, thus permitting streamlined purification of a large number of samples. Robotic protein crystallization screens and optimization utilize both the batch method under oil and vapour diffusion. In order to record and organize the data accumulated by the ISPC, a laboratory information-management system (LIMS) has been developed which facilitates data monitoring and analysis. This permits optimization of conditions at all stages of protein production and structure determination. A set of bioinformatics tools, which are implemented in our LIMS, is utilized to analyze each target.
Albeck
Shira
S
Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Burstein
Yigal
Y
Dym
Orly
O
Jacobovitch
Yossi
Y
Levi
Nurit
N
Meged
Ran
R
Michael
Yigal
Y
Peleg
Yoav
Y
Prilusky
Jaime
J
Schreiber
Gideon
G
Silman
Israel
I
Unger
Tamar
T
Sussman
Joel L
JL
D001331:Automation; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D004247:DNA / Q000737:chemistry; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies; D020407:Internet; D007557:Israel; D008958:Models, Molecular; D010843:Pichia / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry
D011506:Proteins; D011994:Recombinant Proteins; D004247:DNA