Immunochemical approach to the mapping of an assembled epitope of human chorionic gonadotropin: Proximity of CTP-alpha to the receptor binding region of the beta-subunit

A single-step solid-phase RIA (SS-SPRIA) developed in our laboratory using hybridoma culture supernatants has been utilised for the quantitation of epitope-paratope interactions. Using SS-SPRIA as a quantitative tool for the assessment of epitope stability, it was found that several assembled epitopes of human chorionic gonadotropin (hCG) are differentially stable to proteolysis and chemical modification. Based on these observations an approach has been developed for identifying the amino acid residues constituting an epitopic region. This approach has now been used to map an assembled epitope at/near the receptor binding region of the hormone. The mapped site forms a part of the seat belt region and the cystine knot region (C34-C38-C88-C90-H106). The carboxy terminal region of the alpha-subunit forms a part of the epitope indicating its proximity to the receptor binding region. These results are in agreement with the reported receptor binding region identified through other approaches and the X-ray crystal structure of hCG.

Mapping of assembled epitopic regions of human chorionic gonadotropin reveals proximity of CTP alpha to the determinant loop beta 93-100

Three overlapping assembled epitopes of beta hCG have been mapped using MAb probes and a single step solid phase radioimmunoassay. These epitopes have been shown to be at receptor binding region comprising of the loop region beta Cys93-Cys100. Importance of disulphide bonds in maintaining integrity of these epitopes is assessed. Two MAbs (INN 58 and INN 22) interact with the beta region as well as the alpha C-terminal peptide, while the other MAb INN 24 interacts with only the beta region. Cross-reactivity pattern with beta hCG and hLH as web as the reported crystal structure of hCG substantiates the epitope identification. The results demonstrate utility of MAbs as probes in investigations on three-dimensional structure of gonadatropins.

Plasmonic Interactions at Close Proximity in Chiral Geometries: Route toward Broadband Chiroptical Response and Giant Enantiomeric Sensitivity

Chiral metamaterials can have diverse technological applications, such as engineering strongly twisted local electromagnetic fields for sensitive detection of chiral molecules, negative indices of refraction, broadband circular polarization devices, and many more. These are commonly achieved by arranging a group of noble-metal nanoparticles in a chiral geometry, which, for example, can be a helix, whose chiroptical response originates in the dynamic electromagnetic interactions between the localized plasmon modes of the individual nanoparticles. A key question relevant to the chiroptical response of such materials is the role of plasmon interactions as the constituent particles are brought closer, which is investigated in this paper through theoretical and experimental studies. The results of our theoretical analysis, when the particles are brought in close proximity are dramatic, showing a large red shift and enhancement of the spectral width and a near-exponential rise in the strength of the chiroptical response. These predictions were further confirmed with experimental studies of gold and silver nanoparticles arranged on a helical template, where the role of particle separation could be investigated in a systematic manner. The ``optical chirality'' of the electromagnetic fields in the vicinity of the nanoparticles was estimated to be orders of magnitude larger than what could be achieved in all other nanoplasmonic geometries considered so far, implying the suitability of the experimental system for sensitive detection of chiral molecules.

Modelling Proximity Effects in Transverse Magnetic Mode for Lightning Studies

Lightning strike to instrumented and communication towers can be a source of electromagnetic disturbance to the system connected. Long cables running on these towers can get significant induction to their sheath/core, which would then couple to the connected equipments. For a quantitative analysis of the situation, suitable theoretical analysis is necessary. Due to the dominance of the transverse magnetic mode during the fast rising portion of the stroke current, which is the period of significant induction, a full wave solution based on Maxwell's equations is necessary. Owing to the large geometric aspect ratio of tower lattice elements and for feasibility of a numerical solution, the thin-wire formulation for the electric field integral equation is generally adopted. However, the classical thin-wire formulation is not set for handling non-cylindrical conductors like tower lattice elements and the proximity of other conductors. The present work investigates further into a recently proposed method for handling such a situation and optimizes the numerical solution approach.

A mean-reverting stochastic model for the political business cycle

In this article, we look at the political business cycle problem through the lens of uncertainty. The feedback control used by us is the famous NKPC with stochasticity and wage rigidities. We extend the New Keynesian Phillips Curve model to the continuous time stochastic set up with an Ornstein-Uhlenbeck process. We minimize relevant expected quadratic cost by solving the corresponding Hamilton-Jacobi-Bellman equation. The basic intuition of the classical model is qualitatively carried forward in our set up but uncertainty also plays an important role in determining the optimal trajectory of the voter support function. The internal variability of the system acts as a base shifter for the support function in the risk neutral case. The role of uncertainty is even more prominent in the risk averse case where all the shape parameters are directly dependent on variability. Thus, in this case variability controls both the rates of change as well as the base shift parameters. To gain more insight we have also studied the model when the coefficients are time invariant and studied numerical solutions. The close relationship between the unemployment rate and the support function for the incumbent party is highlighted. The role of uncertainty in creating sampling fluctuation in this set up, possibly towards apparently anomalous results, is also explored.

Residue proximity information and protein model discrimination using saturation-suppressor mutagenesis

Identification of residue-residue contacts from primary sequence can be used to guide protein structure prediction. Using Escherichia coli CcdB as the test case, we describe an experimental method termed saturation-suppressor mutagenesis to acquire residue contact information. In this methodology, for each of five inactive CcdB mutants, exhaustive screens for suppressors were performed. Proximal suppressors were accurately discriminated from distal suppressors based on their phenotypes when present as single mutants. Experimentally identified putative proximal pairs formed spatial constraints to recover >98% of native-like models of CcdB from a decoy dataset. Suppressor methodology was also applied to the integral membrane protein, diacylglycerol kinase A where the structures determined by X-ray crystallography and NMR were significantly different. Suppressor as well as sequence co-variation data clearly point to the Xray structure being the functional one adopted in vivo. The methodology is applicable to any macromolecular system for which a convenient phenotypic assay exists.