Differential effects of supplementary affinity tags on the solubility of MBP fusion proteinsJournal of Structural and Functional Genomics - Tập 2 - Trang 83-92 - 2002
Karen M. Routzahn, David S. Waugh
It is difficult to imagine any strategy for high-throughput protein expression
and purification that does not involve genetically engineered affinity tags.
Because of its ability to enhance the solubility and promote the proper folding
of its fusion partners, Escherichia coli maltose-binding protein (MBP) is a
particularly useful affinity tag. However, not all MBP fusion proteins bind
efficiently ... hiện toàn bộ
SUMO fusions and SUMO-specific protease for efficient expression and purification of proteinsJournal of Structural and Functional Genomics - Tập 5 - Trang 75-86 - 2004
Michael P. Malakhov, Michael R. Mattern, Oxana A. Malakhova, Mark Drinker, Stephen D. Weeks, Tauseef R. Butt
SUMO (small ubiquitin-related modifier) modulates protein structure and function
by covalently binding to the lysine side chains of the target proteins. Yeast
cells contain two SUMO proteases, Ulp1 and Ulp2, that cleave sumoylated proteins
in the cell. Ulp1 (SUMO protease 1) processes the SUMO precursor to its mature
form and also de-conjugates SUMO from side chain lysines of target proteins.
Here... hiện toàn bộ
SOn-column Protein Refolding for CrystallizationJournal of Structural and Functional Genomics - Tập 6 - Trang 177-182 - 2005
Natalia Oganesyan, Sung-Hou Kim, Rosalind Kim
One major bottleneck in protein production in Escherichia coli for structural
genomics projects is the formation of insoluble protein aggregates (inclusion
bodies). The efficient refolding of proteins from inclusion bodies is becoming
an important tool that can provide soluble native proteins for structural and
functional studies. Here we report an on-column refolding method established at
the Ber... hiện toàn bộ
Methods and Results for Semi-automated Cloning Using Integrated RoboticsJournal of Structural and Functional Genomics - Tập 6 - Trang 89-94 - 2005
Heath E. Klock, Aprilfawn White, Eric Koesema, Scott A. Lesley
The Joint Center for Structural Genomics (JCSG) has emphasized automation and
parallel processing approaches. Here, we describe automated methods used across
the cloning process with results from JCSG projects. The protocols for PCR,
restriction digests and ligations, as well as for gel electrophoresis and
microtiter plate assays have all been automated. The system has the capacity to
routinely pr... hiện toàn bộ
Project management system for structural and functional proteomics: SesameJournal of Structural and Functional Genomics - Tập 4 - Trang 11-23 - 2003
Zsolt Zolnai, Peter T. Lee, Jing Li, Michael R. Chapman, Craig S. Newman, George N. Phillips, Ivan Rayment, Eldon L. Ulrich, Brian F. Volkman, John L. Markley
A computing infrastructure (Sesame) has been designed to manage and link
individual steps in complex projects. Sesame is being developed to support a
large-scale structural proteomics pilot project. When complete, the system is
expected to manage all steps from target selection to data-bank deposition and
report writing. We report here on the design criteria of the Sesame system and
on results dem... hiện toàn bộ
Heterologous expression of L. major proteins in S. cerevisiae: a test of solubility, purity, and gene recodingJournal of Structural and Functional Genomics - Tập 10 - Trang 233-247 - 2009
Erin Quartley, Andrei Alexandrov, Maryann Mikucki, Frederick S. Buckner, Wim G. Hol, George T. DeTitta, Eric M. Phizicky, Elizabeth J. Grayhack
High level expression of many eukaryotic proteins for structural analysis is
likely to require a eukaryotic host since many proteins are either insoluble or
lack essential post-translational modifications when expressed in E. coli. The
well-studied eukaryote Saccharomyces cerevisiae possesses several attributes of
a good expression host: it is simple and inexpensive to culture, has proven
genetic ... hiện toàn bộ
Erratum to: The NMR solution structures of the five constituent cold-shock domains (CSD) of the human UNR (upstream of N-ras) proteinJournal of Structural and Functional Genomics - Tập 11 Số 2 - Trang 189-189 - 2010
Alexander K. Goroncy, S. Koshiba, N. Tochio, T. Tomizawa, Makoto Inoue, Satoru Watanabe, T. Harada, Akiko Tanaka, Osamu Ohara, T. Kigawa, Shigeyuki Yokoyama
Solution NMR structures of homeodomains from human proteins ALX4, ZHX1, and CASP8AP2 contribute to the structural coverage of the Human Cancer Protein Interaction NetworkJournal of Structural and Functional Genomics - Tập 15 Số 4 - Trang 201-207 - 2014
Xu, Xianzhong, Pulavarti, Surya V. S. R. K., Eletsky, Alexander, Huang, Yuanpeng Janet, Acton, Thomas B., Xiao, Rong, Everett, John K., Montelione, Gaetano T., Szyperski, Thomas
High-quality solution NMR structures of three homeodomains from human proteins
ALX4, ZHX1 and CASP8AP2 were solved. These domains were chosen as targets of a
biomedical theme project pursued by the Northeast Structural Genomics
Consortium. This project focuses on increasing the structural coverage of human
proteins associated with cancer.