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Sialyltransferase is an enzyme that transfers the sialic acid moiety from cytidine-5-monophospho-N-acetylneuraminic acid (CMP-NeuAc) to the carbohydrate group of various glycoproteins. These glycoproteins are involved in inflammation, embryogenesis, immune defence and metastasis of cancer cells by cell-cell interactions or cell-matrix interactions. The alpha-2,6-sialyltransferase PM0188 from Pasteurella multocida was purified using affinity-column chromatographic methods and crystallized using the hanging-drop vapour-diffusion method at 293 K. MAD data were collected to 1.9 A resolution from an SeMet-substituted crystal. The crystal belongs to space group P2(1), with unit-cell parameters a = 52.9, b = 61.0, c = 64.6 A, alpha = gamma = 90, beta = 112.3 degrees. Assuming the presence of one molecule in the asymmetric unit, the solvent content is estimated to be about 45%.

The 4-1BB ligand, a member of the tumour necrosis factor (TNF) family, is an important co-stimulatory molecule that plays a key role in the clonal expansion and survival of CD8+ T cells. Signalling through binding of the 4-1BB ligand and 4-1BB has been reported to enhance CD8+ T-cell expansion and protect activated CD8+ T cells from death. The 4-1BB ligand is an integral protein expressed on activated antigen-presenting cells. The extracellular domain of the 4-1BB ligand fused with glutathione-S-transferase was expressed in Escherichia coli (Origami) and purified by using affinity and ion-exchange column chromatographic methods. Crystals of the 4-1BB ligand were obtained at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected from these crystals to 2.8 A resolution and the crystals belong to space group C2, with unit-cell parameters a = 114.6, b = 73.8, c = 118.50 A, beta = 115.5 degrees.

Purpose: To evaluate the ability of scanning laser polarimetry parameters and a novel deviation map algorithm to discriminate between healthy and early glaucomatous eyes with localized visual field (VF) defects confined to one hemifield., Design: Prospective case-control study., Participants: Seventy glaucomatous eyes with localized VF defects and 66 normal controls., Methods: A Humphrey field analyzer 24-2 full-threshold test and scanning laser polarimetry with variable corneal compensation were used., Main Outcome Measures: We assessed the sensitivity and specificity of scanning laser polarimetry parameters, sensitivity and cutoff values for scanning laser polarimetry deviation map algorithms at different specificity values (80%, 90%, and 95%) in the detection of glaucoma, and correlations between the algorithms of scanning laser polarimetry and of the pattern deviation derived from Humphrey field analyzer testing., Results: There were significant differences between the glaucoma group and normal subjects in the mean parametric values of the temporal, superior, nasal, inferior, temporal (TSNIT) average, superior average, inferior average, and TSNIT standard deviation (SD) (P<0.05). The sensitivity and specificity of each scanning laser polarimetry variable was as follows: TSNIT, 44.3% (95% confidence interval [CI], 39.8%-49.8%) and 100% (95.4%-100%); superior average, 30% (25.5%-34.5%) and 97% (93.5%-100%); inferior average, 45.7% (42.2%-49.2%) and 100% (95.8%-100%); and TSNIT SD, 30% (25.9%-34.1%) and 97% (93.2%-100%), respectively (when abnormal was defined as P<0.05). Based on nerve fiber indicator cutoff values of > or =30 and > or =51 to indicate glaucoma, sensitivities were 54.3% (50.1%-58.5%) and 10% (6.4%-13.6%), and specificities were 97% (93.2%-100%) and 100% (95.8%-100%), respectively. The range of areas under the receiver operating characteristic curves using the scanning laser polarimetry deviation map algorithm was 0.790 to 0.879. Overall sensitivities combining each probability scale and severity score at 80%, 90%, and 95% specificities were 90.0% (95% CI, 86.4%-93.6%), 71.4% (67.4%-75.4%), and 60.0% (56.2%-63.8%), respectively. There was a statistically significant correlation between the scanning laser polarimetry severity score and the VF severity score (R2 = 0.360, P<0.001)., Conclusions: Scanning laser polarimetry parameters may not be sufficiently sensitive to detect glaucomatous patients with localized VF damage. Our algorithm using the scanning laser polarimetry deviation map may enhance the understanding of scanning laser polarimetry printouts in terms of the locality, deviation size, and severity of localized retinal nerve fiber layer defects in eyes with localized VF loss.

The aim of this study was to evaluate the efficacy of 0.005% latanoprost in lowering intraocular pressure (IOP) in patients with chronic angle-closure glaucoma (CACG) and no visible ciliary-body face. Fourteen eyes of 14 Korean patients with CACG with 360 degrees of peripheral anterior synechiae (PAS) and an IOP greater than 21 mmHg without medication were treated with 0.005% latanoprost once-daily. All patients completed 3 months of treatment with latanoprost. The IOP, which was 30.3 +/- 4.5 (mean +/- standard deviation) mmHg at baseline, decreased to 22.6 +/- 4.9 mmHg after 1 week, 19.6 +/- 5.5 mmHg after 1 month, 19.4 +/- 4.9 mmHg after 2 months, and 21.5 +/- 5.9 mmHg after 3 months of treatment with latanoprost (P < 0.01 for each). Ultrasound biomicroscopy of the anterior chamber angle showed anterior bowing of the iris with total occlusion of the angle by PAS, except for 5 eyes with focal microscopic openings to the ciliary-body face at various angles. Adverse ocular events were well-tolerated and transient. In this preliminary study, treatment with 0.005% latanoprost once-daily resulted in a significant reduction in IOP in CACG patients with 360 degrees of PAS on gonioscopy. Our results suggest that latanoprost may be considered as a therapy of choice in these rare cases.

Recent crystallographic studies have provided significant new insight into how receptor tyrosine kinases from the EGF receptor or ErbB family are regulated by their growth factor ligands. EGF receptor dimerization is mediated by a unique dimerization arm, which becomes exposed only after a dramatic domain rearrangement is promoted by growth factor binding. ErbB2, a family member that has no ligand, has its dimerization arm constitutively exposed, and this explains several of its unique properties. We outline a mechanistic view of ErbB receptor homo- and heterodimerization, which suggests new approaches for interfering with these processes when they are implicated in human cancers.

HER2 (also known as Neu, ErbB2) is a member of the epidermal growth factor receptor (EGFR; also known as ErbB) family of receptor tyrosine kinases, which in humans includes HER1 (EGFR, ERBB1), HER2, HER3 (ERBB3) and HER4 (ERBB4). ErbB receptors are essential mediators of cell proliferation and differentiation in the developing embryo and in adult tissues, and their inappropriate activation is associated with the development and severity of many cancers. Overexpression of HER2 is found in 20-30% of human breast cancers, and correlates with more aggressive tumours and a poorer prognosis. Anticancer therapies targeting ErbB receptors have shown promise, and a monoclonal antibody against HER2, Herceptin (also known as trastuzumab), is currently in use as a treatment for breast cancer. Here we report crystal structures of the entire extracellular regions of rat HER2 at 2.4 A and human HER2 complexed with the Herceptin antigen-binding fragment (Fab) at 2.5 A. These structures reveal a fixed conformation for HER2 that resembles a ligand-activated state, and show HER2 poised to interact with other ErbB receptors in the absence of direct ligand binding. Herceptin binds to the juxtamembrane region of HER2, identifying this site as a target for anticancer therapies.

Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. To activate the receptor, EGF binding must promote a large domain rearrangement that exposes this dimerization interface. This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists.

We have determined the 2.6 angstrom crystal structure of the entire extracellular region of human HER3 (ErbB3), a member of the epidermal growth factor receptor (EGFR) family. The structure consists of four domains with structural homology to domains found in the type I insulin-like growth factor receptor. The HER3 structure reveals a contact between domains II and IV that constrains the relative orientations of ligand-binding domains and provides a structural basis for understanding both multiple-affinity forms of EGFRs and conformational changes induced in the receptor by ligand binding during signaling. These results also suggest new therapeutic approaches to modulating the behavior of members of the EGFR family.

Over 20 enzymes denoted as cyclomaltodextrinase, maltogenic amylase, or neopullulanase that share 40-86% sequence identity with each other are found in public data bases. These enzymes are distinguished from typical alpha-amylases by containing a novel N-terminal domain and exhibiting preferential substrate specificities for cyclomaltodextrins (CDs) over starch. In this research field, a great deal of confusion exists regarding the features distinguishing the three groups of enzymes from one another. Although a different enzyme code has been assigned to each of the three different enzyme names, even a single differentiating enzymatic property has not been documented in the literature. On the other hand, an outstanding question related to this issue concerns the structural basis for the preference of these enzymes for CDs. To clarify the confusion and to address this question, we have determined the structures of two enzymes, one from alkalophilic Bacillus sp. I-5 and named cyclomaltodextrinase and the other from a Thermus species and named maltogenic amylase. The structure of the Bacillus enzyme reveals a dodecameric assembly composed of six copies of the dimer, which is the structural and functional unit of the Thermus enzyme and an enzyme named neopullulanase. The structure of the Thermus enzyme in complex with beta-CD led to the conclusion that Trp47, a well conserved N-terminal domain residue, contributes greatly to the preference for beta-CD. The common dimer formation through the novel N-terminal domain, which contributes to the preference for CDs by lining the active-site cavity, convincingly indicates that the three groups of enzymes are not different enough to preserve the different names and enzyme codes.