Item 1: October 2004 Publications by Members:

1:  Swaminathan CP, Wais N, Vyas VV, Velikovsky CA, Moretta A, Moretta L, Biassoni R, Mariuzza RA, Dimasi N.
Entropically Assisted Carbohydrate Recognition by a Natural Killer Cell-Surface Receptor.
Chembiochem. 2004 Oct 29;5(11):1571-1575 PMID: 15515091

2:  Canagarajah B, Leskow FC, Ho JY, Mischak H, Saidi LF, Kazanietz MG, Hurley JH.
Structural Mechanism for Lipid Activation of the Rac-Specific GAP,
beta2-Chimaerin.
Cell. 2004 Oct 29;119(3):407-18. PMID: 15507211

3:  Saxena AK, Singh K, Long CA, Garboczi DN.
Preparation, crystallization and preliminary X-ray analysis of a complex
between the Plasmodium vivax sexual stage 25 kDa protein Pvs25 and a malaria transmission-blocking antibody Fab fragment.
Acta Crystallogr D Biol Crystallogr. 2004 Nov;60(Pt 11):2054-7. Epub 2004 Oct 20. PMID: 15502325

4:  Schubot FD, Waugh DS.
A pivotal role for reductive methylation in the de novo crystallization of a
ternary complex composed of Yersinia pestis virulence factors YopN, SycN and YscB.
Acta Crystallogr D Biol Crystallogr. 2004 Nov;60(Pt 11):1981-6. PMID: 15502305

5:  Zwart PH, Banumathi S, Dauter M, Dauter Z.
Radiation-damage-induced phasing with anomalous scattering: substructure solution and phasing.
Acta Crystallogr D Biol Crystallogr. 2004 Nov;60(Pt 11):1958-63. PMID: 15502302

6:  Yamamoto Y, Moore R, Goldsworthy TL, Negishi M, Maronpot RR.
The orphan nuclear receptor constitutive active/androstane receptor is
essential for liver tumor promotion by phenobarbital in mice.
Cancer Res. 2004 Oct 15;64(20):7197-200. PMID: 15492232

7:  Fritz TA, Hurley JH, Trinh LB, Shiloach J, Tabak LA.
The beginnings of mucin biosynthesis: the crystal structure of
UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferase-T1.
Proc Natl Acad Sci U S A. 2004 Oct 26;101(43):15307-12. PMID: 15486088

8:  Guarne A, Ramon-Maiques S, Wolff EM, Ghirlando R, Hu X, Miller JH, Yang W.
Structure of the MutL C-terminal domain: a model of intact MutL and its roles in mismatch repair.
EMBO J. 2004 Oct 27;23(21):4134-45. PMID: 15470502

9:  Dauter Z.
Phasing in iodine for structure determination.
Nat Biotechnol. 2004 Oct;22(10):1239-40. PMID: 15470459

10:  Iwahara J, Schwieters CD, Clore GM.
Characterization of nonspecific protein-DNA interactions by 1H paramagnetic relaxation enhancement.
J Am Chem Soc. 2004 Oct 13;126(40):12800-8. PMID: 15469275

11:  Agniswamy J, Lei B, Musser JM, Sun PD.
Insight of host immune evasion mediated by two variants of group a
streptococcus Mac protein.
J Biol Chem. 2004 Oct 4. PMID: 15466462

12:  Liang X, Young JJ, Boone SA, Waugh DS, Duesbery NS.
Involvement of domain II in toxicity of anthrax lethal factor.
J Biol Chem. 2004 Oct 1. PMID: 15465830

13:  Ramakrishnan B, Boeggeman E, Ramasamy V, Qasba PK.
Structure and catalytic cycle of beta-1,4-galactosyltransferase.
Curr Opin Struct Biol. 2004 Oct;14(5):593-600. PMID: 15465321

14:  Janda I, Devedjiev Y, Derewenda U, Dauter Z, Bielnicki J, Cooper DR, Graf PC, Joachimiak A, Jakob U, Derewenda ZS.
The crystal structure of the reduced, Zn2+-bound form of the B. subtilis Hsp33 chaperone and its implications for the activation mechanism.
Structure (Camb). 2004 Oct;12(10):1901-7. PMID: 15458638

15:  Schalk IJ, Yue WW, Buchanan SK.
Recognition of iron-free siderophores by TonB-dependent iron transporters.
Mol Microbiol. 2004 Oct;54(1):14-22. PMID: 15458401

16:  Ramakrishnan B, Boeggeman E, Qasba PK.
 Effect of the Met344His mutation on the conformational dynamics of bovine beta-1,4-galactosyltransferase: crystal structure of the Met344His mutant in complex with chitobiose.
Biochemistry. 2004 Oct 5;43(39):12513-22. PMID: 15449940

17:  Ofek G, Tang M, Sambor A, Katinger H, Mascola JR, Wyatt R, Kwong PD.
Structure and mechanistic analysis of the anti-human immunodeficiency virus type 1 antibody 2F5 in complex with its gp41 epitope.
J Virol. 2004 Oct;78(19):10724-37. PMID: 15367639

18:  Zou Z, Sun PD.
Overexpression of human transforming growth factor-beta1 using a recombinant CHO cell expression system.
Protein Expr Purif. 2004 Oct;37(2):265-72. PMID: 15358346

19:  Moon AF, Edavettal SC, Krahn JM, Munoz EM, Negishi M, Linhardt RJ, Liu J, Pedersen LC.
Structural analysis of the sulfotransferase (3-o-sulfotransferase isoform 3)
involved in the biosynthesis of an entry receptor for herpes simplex virus 1.
J Biol Chem. 2004 Oct 22;279(43):45185-93. PMID: 15304505

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.

Recommended Readings:

(1) Anomalous-scatterer-mediated crystal-packing interactions

(2) Strategies in making cross-linked enzyme crystals

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.