Item 1: August 2004 Publications by Members:

1:  Hierro A, Sun J, Rusnak AS, Kim J, Prag G, Emr SD, Hurley JH.
Structure of the ESCRT-II endosomal trafficking complex.
Nature. 2004 Aug 25, PMID: 15329733

2:  Chang C, Pluckthun A, Wlodawer A.
Crystal structure of a truncated version of the phage lambda protein gpD.
Proteins. 2004 Aug 12 , PMID: 15317025

3:  Clore GM, Schwieters CD.
Amplitudes of Protein Backbone Dynamics and Correlated Motions in a Small alpha/beta Protein: Correspondence of Dipolar Coupling and Heteronuclear Relaxation Measurements.
Biochemistry. 2004 Aug 24;43(33):10678-10691. PMID: 15311929

4:  Ronning DR, Li Y, Perez ZN, Ross PD, Hickman AB, Craig NL, Dyda F.
The carboxy-terminal portion of TnsC activates the Tn7 transposase through a specific interaction with TnsA.
EMBO J. 2004 Aug 4;23(15):2972-81. Epub 2004 Jul 15. PMID: 15257292

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.