Blockade of Neuronal α7-nAChR by α-Conotoxin ImI Explained by Computational Scanning and Energy Calculations

α-Conotoxins potently inhibit isoforms of nicotinic acetylcholine receptors (nAChRs), which are essential for neuronal and neuromuscular transmission. They are also used as neurochemical tools to study nAChR physiology and are being evaluated as drug leads to treat various neuronal disorders. A number of experimental studies have been performed to investigate the structure-activity relationships of conotoxin/nAChR complexes. However, the structural determinants of their binding interactions are still ambiguous in the absence of experimental structures of conotoxin-receptor complexes. In this study, the binding modes of α-conotoxin ImI to the α7-nAChR, currently the best-studied system experimentally, were investigated using comparative modeling and molecular dynamics simulations. The structures of more than 30 single point mutants of either the conotoxin or the receptor were modeled and analyzed. The models were used to explain qualitatively the change of affinities measured experimentally, including some nAChR positions located outside the binding site. Mutational energies were calculated using different methods that combine a conformational refinement procedure (minimization with a distance dependent dielectric constant or explicit water, or molecular dynamics using five restraint strategies) and a binding energy function (MM-GB/SA or MM-PB/SA). The protocol using explicit water energy minimization and MM-GB/SA gave the best correlations with experimental binding affinities, with an R2 value of 0.74. The van der Waals and non-polar desolvation components were found to be the main driving force for binding of the conotoxin to the nAChR. The electrostatic component was responsible for the selectivity of the various ImI mutants. Overall, this study provides novel insights into the binding mechanism of α-conotoxins to nAChRs and the methodological developments reported here open avenues for computational scanning studies of a rapidly expanding range of wild-type and chemically modified α-conotoxins

Penilaian Kinerja di Divisi Pelayanan Terminal untuk Mengukur Keberhasilan Organisasi di PT. Pelabuhan Indonesia I Belawan

PT Pelabuhan Indonesia I merupakan Perusahaan yang bergerak dibidang penyelenggaraan dan pengusahaan jasa kepelabuhanan bertujuan mengoptimalisasi pemanfaatan sumber daya yang dimiliki Perusahaan untuk menghasilkan barang atau jasa yang bermutu tinggi dan berdaya saing kuat maka kebutuhan akan tingkat kinerja yang tinggi dari setiap karyawan merupakan hal yang mutlak bagi PT. Pelabuhan Indonesia I. Berdasarkan hasil penilaian persentase kompetensi dan produktivitas yang telah ada, persentase realisasi yang dicapai lebih kecil dari angka target yang ditetapkan oleh Perusahaan. kesenjangan ini mengindikasikan bahwa kinerja karyawan belum maksimal dan perlu adanya penilaian kinerja lebih lanjut. Penilaian kinerja merupakan suatu evaluasi hasil kerja dari seorang karyawan secara sistematis yang berhubungan dengan jabatannya dan potensi yang dimilikinya untuk dikembangkan. Penilaian kinerja ini diharapkan dapat menjadi umpan Balik bagi karyawan tersebut tentang prestasi kerjanya selama ini dan mengembangkan kemampuannya lebih lanjut. Penilaian kinerja bertujuan untuk akan meningkatkan prestasi organisasi. Penelitian ini menggunakan kuesioner, software SPSS 17.0 dan Matlab sebagai instrumen penelitian. Hasil perhitungan menunjukkan bahwa pada level top manajemen, Manajer pelayanan terminal memiliki penilaian kinerja tertinggi dengan nilai eigen 2.1903. Pada level middle manajemen, Supervisor Pelayanan Operasi Senior memiliki penilaian kinerja tertinggi dengan nilai eigen 2.7424. Pada level lower manajemen, penilaian kinerja tertinggi adalah Pelaksana Perencanaan & Pengendalian Operasi Senior dengan nilai eigen 2.6141. Jabatan Asisten Manajer Pelayanan Operasi dan Jabatan Pelaksana Pelayanan Operasi Senior merupakan jabatan yang memiliki penilaian kinerja yang sangat rendah sehingga memerlukan evaluasi lebih lanjut. Hal ini dapat dilihat dari nilai eigen kedua jabatan yang bernilai negatif, masing-masing yaitu -0.0005 dan -0.000

Dual-targeting anti-angiogenic cyclic peptides as potential drug leads for cancer therapy

Peptide analogues derived from bioactive hormones such as somatostatin or certain growth factors have great potential as angiogenesis inhibitors for cancer applications. In an attempt to combat emerging drug resistance many FDA-approved anti-angiogenesis therapies are co-administered with cytotoxic drugs as a combination therapy to target multiple signaling pathways of cancers. However, cancer therapies often encounter limiting factors such as high toxicities and side effects. Here, we combined two anti-angiogenic epitopes that act on different pathways of angiogenesis into a single non-toxic cyclic peptide framework, namely MCoTI-II (Momordica cochinchinensis trypsin inhibitor-II), and subsequently assessed the anti-angiogenic activity of the novel compound. We hypothesized that the combination of these two epitopes would elicit a synergistic effect by targeting different angiogenesis pathways and result in improved potency, compared to that of a single epitope. This novel approach has resulted in the development of a potent, non-toxic, stable and cyclic analogue with nanomolar potency inhibition in in vitro endothelial cell migration and in vivo chorioallantoic membrane angiogenesis assays. This is the first report to use the MCoTI-II framework to develop a 2-in-1 anti-angiogenic peptide, which has the potential to be used as a form of combination therapy for targeting a wide range of cancers

Dissecting the oxidative folding of circular cystine knot miniproteins

Cyclotides are plant proteins with exceptional stability owing to the presence of a cyclic backbone and three disulfide bonds arranged in a cystine knot motif. Accordingly, they have been proposed as templates to stabilize bioactive epitopes in drug-design applications. The two main subfamilies, referred to as the Möbius and bracelet cyclotides, require dramatically different in vitro folding conditions to achieve the native fold. To determine the underlying elements that influence cyclotide folding, we examined the in vitro folding of a suite of hybrid cyclotides based on combination of the Möbius cyclotide kalata B1 and the bracelet cyclotide cycloviolacin O1. The folding pathways of the two cyclotide subfamilies were found to be different and influenced by specific residues within intercysteine loops 2 and 6. Two changes in these loops, a substitution in loop 2 and an addition in loop 6, enabled the folding of a cycloviolacin O1 analogue under conditions in which folding does not occur in vitro for the native peptide. A key intermediate contains a native-like hairpin structure that appears to be a nucleation locus early in the folding process. Overall, these mechanistic findings on the folding of cyclotides are potentially valuable for the design of new drug leads. Copyright Mary Ann Liebert, Inc

ConoServer: updated content, knowledge, and discovery tools in the conopeptide database

ConoServer (http://www.conoserver.org) is a database specializing in the sequences and structures of conopeptides, which are toxins expressed by marine cone snails. Cone snails are carnivorous gastropods, which hunt their prey using a cocktail of toxins that potently subvert nervous system function. The ability of these toxins to specifically target receptors, channels and transporters of the nervous system has attracted considerable interest for their use in physiological research and as drug leads. Since the founding publication on ConoServer in 2008, the number of entries in the database has nearly doubled, the interface has been redesigned and new annotations have been added, including a more detailed description of cone snail species, biological activity measurements and information regarding the identification of each sequence. Automatically updated statistics on classification schemes, three-dimensional structures, conopeptide-bearing species and endoplasmic reticulum signal sequence conservation trends, provide a convenient overview of current knowledge on conopeptides. Transcriptomics and proteomics have began generating massive numbers of new conopeptide sequences, and two dedicated tools have been recently implemented in ConoServer to standardize the analysis of conopeptide precursor sequences and to help in the identification by mass spectrometry of toxins whose sequences were predicted at the nucleic acid level

Proteome analysis of the hyaluronic acid-producing bacterium, Streptococcus zooepidemicus

Background: Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a commensal of horses and an opportunistic pathogen in many animals and humans. Some strains produce copious amounts of hyaluronic acid, making S. zooepidemicus an important industrial microorganism for the production of this valuable biopolymer used in the pharmaceutical and cosmetic industry. Encapsulation by hyaluronic acid is considered an important virulence factor in other streptococci, though the importance in S. zooepidemicus remains poorly understood. Proteomics may provide a better understanding of virulence factors in S. zooepidemicus, facilitate the design of better diagnostics and treatments, and guide engineering of superior production strains