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Journal Issue - Volume 16 Issue 12 (December 2007)

Abstract Pathogens have evolved numerous strategies to infect their hosts, while hosts have evolved immune responses and other defenses to these foreign challenges. The vast majority of host–pathogen interactions involve protein–protein recognition, yet our current understanding of these interactions is limited. Here, we present and apply a computational whole‐genome protocol that generates testable predictions of host–pathogen...

Abstract α‐Actinin is an actin bundling protein that regulates cell adhesion by directly linking actin filaments to integrin adhesion receptors. Phosphatidylinositol (4,5)‐diphosphate (PtdIns (4,5)‐P2) and phosphatidylinositol (3,4,5)‐triphosphate (PtdIns (3,4,5)‐P3) bind to the calponin homology 2 domain of α‐actinin, regulating its interactions with actin filaments and integrin receptors. In this study, we examine the mechanism by which...

Abstract The extracellular adherence protein (Eap) of Staphylococcus aureus participates in a wide range of protein–protein interactions that facilitate the initiation and dissemination of Staphylococcal disease. In this report, we describe the use of a multidisciplinary approach to characterize the solution structure of full‐length Eap. In contrast to previous reports suggesting that a six‐domain isoform of Eap undergoes multimerization, sedimentation ...

Abstract Peptidyl‐prolyl isomerase (PPIase) activity is exhibited by many proteins belonging to the PPIase family. However, the catalytic mechanism of this activity remains to be completely elucidated. Here, we selected human FK506‐binding protein 12 (FKBP12) as the model PPIase and investigated the nature of amino acid residues essential for the activity. The crystal structures of several complexes of PPIase with short peptides...

Abstract The p90 ribosomal S6 kinases (RSKs) also known as MAPKAP‐Ks are serine/threonine protein kinases that are activated by ERK or PDK1 and act as downstream effectors of mitogen‐activated protein kinase (MAPK). RSK1, a member of the RSK family, contains two distinct kinase domains in a single polypeptide chain, the regulatory C‐terminal kinase domain (CTKD) and the catalytic N‐terminal kinase domain (NTKD). Autophosphorylation...

Abstract In class C β‐lactamases, the strictly conserved Asn152 forms part of an extended active‐site hydrogen‐bonding network. To probe its role in catalysis, all 19 mutants of Enterobacter cloacae P99 cephalosporinase Asn152 were simultaneously constructed and screened in Escherichia coli for their in vivo activity. The screen identified the previously uncharacterized mutants Asn152Ser, Asn152Thr, and Asn152Gly, which possess significant...

Abstract Two high‐resolution structures of a double mutant of bacterial cholesterol oxidase in the presence or absence of a ligand, glycerol, are presented, showing the trajectory of glycerol as it binds in a Michaelis complex‐like position in the active site. A group of three aromatic residues forces the oxidized isoalloxazine moiety to bend along the N5‐N10 axis as a response to the binding of glycerol in the active site. Movement...

Abstract N‐Acetylglucosamine‐1‐phosphate uridyltransferase (GlmU) catalyzes the first step in peptidoglycan biosynthesis in both Gram‐positive and Gram‐negative bacteria. The products of the GlmU reaction are essential for bacterial survival, making this enzyme an attractive target for antibiotic drug discovery. A series of Haemophilus influenzae GlmU (hiGlmU) structures were determined by X‐ray crystallography in order to provide structural...

Abstract The efficiency of Helicobacter pylori as a mucosal pathogen is caused by unique soluble and integral membrane proteins, which allow its survival at acidic pH and successful colonization of the gastric environment. With about one‐fourth of the H. pylori's proteome comprising integral membrane proteins, the need for solution of their three‐dimensional (3D) structures becomes persistent as it can potentially drive the generation of more...

Abstract The translocated intimin receptor (TIR) of enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC) is required for EPEC and EHEC infections, which cause widespread illness across the globe. TIR is translocated via a type‐III secretion system into the intestinal epithelial cell membrane, where it serves as an anchor for E. coli attachment via its binding partner intimin. While many aspects of EPEC and EHEC infection are now well...

Abstract Assembly of the adenovirus (Ad) homotrimeric fiber protein is nucleated by its C‐terminal knob domain, which itself can trimerize when expressed as a recombinant protein fragment. The non‐interlocked, globular structure of subunits in the knob trimer implies that trimers assemble from prefolded monomers through a dimer intermediate, but these intermediates have not been observed and the mechanism of assembly therefore...

Abstract Computational design of surface charge–charge interactions has been demonstrated to be an effective way to increase both the thermostability and the stability of proteins. To test the robustness of this approach for proteins with predominantly β‐sheet secondary structure, the chicken isoform of the Fyn SH3 domain was used as a model system. Computational analysis of the optimal distribution of surface charges showed that...

Abstract Wild‐type green fluorescent protein (wt‐GFP) has a prominent absorbance band centered at ∼395 nm, attributed to the neutral chromophore form. The green emission arising upon excitation of this band results from excited‐state proton transfer (ESPT) from the chromophore hydroxyl, through a hydrogen‐bond network proposed to consist of a water molecule and Ser205, to Glu222. Although evidence for Glu222 as a terminal proton...

Abstract Phosphorylation plays an important role in regulation of protein kinase C delta (PKCδ). To date, three Ser/Thr residues (Thr 505, Ser 643, and Ser 662) and nine tyrosine residues (Tyr 52, Tyr 64, Tyr 155, Tyr 187, Tyr 311, Tyr 332, Tyr 512, Tyr 523, and Tyr 565) have been defined as regulatory phosphorylation sites for this protein (rat PKCδ numbering). We combined doxycycline‐regulated inducible gene expression technology...

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