Item 1: September 2004 Publications by Members:

1:  Miller GJ, Hurley JH. "Crystal structure of the catalytic core of inositol 1,4,5-trisphosphate 3-kinase." Mol Cell. 2004 Sep 10;15(5):703-11.
PMID: 15350215

2:  Squires EJ, Sueyoshi T, Negishi M. "Cytoplasmic localization of PXR and ligand-dependent nuclear translocation in mouse liver." J Biol Chem. 2004 Sep 2 PMID: 15347657

3:  Vidal AE, Kannouche PP, Podust VN, Yang W, Lehmann AR, Woodgate R. "PCNA-dependent coordination of the biological functions of human DNA polymerase." J Biol Chem. 2004 Sep 1. PMID: 15342632

4:  LaRonde-LeBlanc N, Wlodawer A. "Crystal structure of A. fulgidus Rio2 defines a new family of serine protein kinases." Structure (Camb). 2004 Sep;12(9):1585-94. PMID: 15341724

5:  Kodama S, Koike C, Negishi M, Yamamoto Y. "Nuclear receptors CAR and PXR cross talk with FOXO1 to regulate genes that encode drug-metabolizing and gluconeogenic enzymes." Mol Cell Biol. 2004 Sep;24(18):7931-40. PMID: 15340055

6:  Phan J, Tropea JE, Waugh DS. "Structure of the Yersinia pestis type III secretion chaperone SycH in complex with a stable fragment of YscM2." Acta Crystallogr D Biol Crystallogr. 2004 Sep;60(Pt 9):1591-9. PMID: 15333930

7:  Sekar K, Rajakannan V, Velmurugan D, Yamane T, Thirumurugan R, Dauter M, Dauter Z. "A redetermination of the structure of the triple mutant (K53,56,120M) of phospholipase A2 at 1.6 A resolution using sulfur-SAS at 1.54 A wavelength." Acta Crystallogr D Biol Crystallogr. 2004 Sep;60(Pt 9):1586-90. PMID: 15333929

8:  Hierro A, Sun J, Rusnak AS, Kim J, Prag G, Emr SD, Hurley JH. "Structure of the ESCRT-II endosomal trafficking complex." Nature. 2004 Sep 9;431(7005):221-5. PMID: 15329733

9:  Miclet E, Williams Jr DC, Clore GM, Bryce DL, Boisbouvier J, Bax A. "Relaxation-optimized NMR spectroscopy of methylene groups in proteins and nucleic acids." J Am Chem Soc. 2004 Sep 1;126(34):10560-70.
PMID: 15327312

10:  Legler PM, Cai M, Peterkofsky A, Clore GM. "Three-dimensional solution structure of the cytoplasmic B domain of the mannitol transporter IImannitol of the Escherichia coli phosphotransferase system." J Biol Chem. 2004 Sep 10;279(37):39115-21. PMID: 15258141

11:  Swietnicki W, O'Brien S, Holman K, Cherry S, Brueggemann E, Tropea JE, Hines HB, Waugh DS, Ulrich RG. "Novel protein-protein interactions of the Yersinia pestis type III secretion system elucidated with a matrix analysis by surface plasmon resonance and mass spectrometry."
J Biol Chem. 2004 Sep 10;279(37):38693-700. PMID: 15213222

Item 2: Tips and Tricks in Crystallography

This section is always open for contributions. Click for Introduction and tips and tricks in Crystallization, Derivatization, Diffraction, Symmetry, Structure Solution, Structure Refinement, and Structure Analysis.

Item 3: Topic Discussion - Protein Expression and High-throughput Expression Systems

Protein Structure Initiative Centers of NIH/NIGMS: Better Tools and Knowledge for Macromolecular Structure Determination.

Dr. Dominic Esposito (Protein Expression Laboratory, NCI-Frederick): Parallel Protein Expression Strategies for Structural Biology

The genomics era has opened up an overwhelming number of possibilities for structural biologists, with tens of thousands of new proteins waiting to be explored. The most interesting of these are human genes and closely related homologs which encode proteins involved in various aspects of human disease. Unfortunately, the biggest bottleneck in exploring this large protein space lies at the level of protein expression. While most prokaryotic genes are readily expressed in soluble form in Escherichia coli, many genes from eukaryotes, particularly those from humans, are very difficult to express in native form in heterogonous organisms. Solving the problem of expressing soluble proteins in purity and quantity enough for structural biology is a pressing concern being addressed these days in nearly every major university, government, and industrial setting.

At the NCI-Frederick Protein Expression Laboratory (PEL), we have taken the approach of developing highly parallel methods for protein expression.  Unlike genomic techniques such as cloning, protein expression rarely offers any consistency from experiment to experiment.  Each protein is its own unique molecule, and must be handled as such; we can draw conclusions from various experimental parameters, but to this point, we are unable to accurately predict ahead of time how a particular protein will behave. For that reason, we view a parallel approach as the most time and cost efficient way to deal with individual proteins of interest.

Click for previous discussions on: NCS, Missing Atoms, Trends in Crystallography, and Absorption Correction.