Partially native intermediates mediate misfolding of SOD1 in single-molecule folding trajectories.

Prion-like misfolding of superoxide dismutase 1 (SOD1) is associated with the disease ALS, but the mechanism of misfolding remains unclear, partly because misfolding is difficult to observe directly. Here we study the most misfolding-prone form of SOD1, reduced un-metallated monomers, using optical tweezers to measure unfolding and refolding of single molecules. We find that the folding is more complex than suspected, resolving numerous previously undetected intermediate states consistent with the formation of individual β-strands in the native structure.

Photostable Copper Nanoclusters: Compatible Förster Resonance Energy-Transfer Assays and a Nanothermometer.

To address the concern of material chemists over the issue of stability and photoluminescent (PL) characteristics of Cu nanoclusters (NCs), herein we present an efficient protocol discussing PL Cu NCs (Cu/HSA) having blue emission and high photostability. These PL NCs were illustrated as efficient probes for Förster resonance energy transfer (FRET) with a compatible fluorophore (Coumarin 153). Our spectroscopic results were well complemented by our molecular docking calculations, which also favored our proposed mechanism for Cu NC formation.

Structural aspects of a protein-surfactant assembly: native and reduced States of human serum albumin.

The inherently present seventeen disulfide bonds of the circulatory protein, human serum albumin (HSA) provide the necessary structural stability. Various spectroscopic approaches were used to investigate the effect of reduction of these disulfide bonds and its binding with the anionic surfactant, sodium dodecyl sulfate (SDS). Based on several spectroscopic analyses, our investigations highlight the following interesting aspects: (1) HSA on reduction loses not only its tertiary structure but also a significant amount of secondary structure as well.

Temperature induced morphological transitions from native to unfolded aggregated States of human serum albumin.

The circulatory protein, human serum albumin (HSA), is known to have two melting point temperatures, 56 and 62 °C. In this present manuscript, we investigate the interaction of HSA with a synthesized bioactive molecule 3-pyrazolyl 2-pyrazoline (PZ). The sole tryptophan amino acid residue (Trp214) of HSA and PZ forms an excellent FRET pair and has been used to monitor the conformational dynamics in HSA as a function of temperature.

Luminescent silver nanoclusters acting as a label-free photoswitch in metal ion sensing.

In this work, we report the application of protein-templated Ag nanoclusters as a luminescent photoswitch for the detection of metal ions. Ag nanoclusters were synthesized using the circulatory protein human serum albumin (HSA) as a template, whose synthetic procedure can be tuned to make them toggle between blue-emitting (Ag9:HSA) and red-emitting (Ag14:HSA) nanoclusters. The photoluminescence (PL) intensity of Ag9:HSA was quenched significantly in the presence of 1 mM Co(II) ions.

Microheterogeneity and microviscosity of F127 micelle: the counter effects of urea and temperature.

F127 is the most widely studied triblock copolymer and due to the presence of very long polypropylene oxide (PPO) and polyethylene oxide (PEO) groups, F127 micelle has different microenvironments clearly separated into core, corona, and peripheral regions. Urea has been known to have adverse effects on the micellar properties and causes demicellization and solvation; on the other hand, rise in temperature causes micellization and solvent evacuation from the core and corona regions. In the present study, we have investigated the microheterogeneity of the core, corona, and peripheral regions of the F127 micelle using red edge excitation shift (REES) at different temperatures and urea concentrations and correlated the effect of both on the micellar system.

Investigating the evolution of drug mediated silver nanoparticles.

Silver nanoparticles have been prepared by a one-pot synthetic technique using the antibacterial drug, Ciprofloxacin (Cp), under optimized conditions. Several techniques are used to characterize the free and nanoparticle bound states of Cp. The time dependent release of the drug molecules from the nanoparticles proves its importance in drug delivery.

Binding, unfolding and refolding dynamics of serum albumins.

Background: The serum albumins (human and bovine serum albumin) occupy a seminal position among all proteins investigated until today as they are the most abundant circulatory proteins. They play the major role of a transporter of many bio-active substances which include various fatty acids, drug molecules, and amino acids to the target cells. Hence, studying the interaction of these serum albumins with different binding agents has attracted enormous research interests from decades.

Reversibility in protein folding: effect of β-cyclodextrin on bovine serum albumin unfolded by sodium dodecyl sulphate.

The mechanism by which the protein bovine serum albumin undergoes unfolding induced by the anionic surfactant sodium dodecyl sulphate (SDS) and then the subsequent refolding brought in by β-Cyclodextrin (β-CD) was studied by steady-state fluorescence, time resolved measurements and Circular Dichroism (CD) spectroscopy. The prominent findings of this investigation are (i) SDS unfolds the protein in a sequential manner passing through three different phases of binding of SDS followed by a saturation phase; (ii) the refolding process is initiated through inclusion/removal of SDS molecules by β-CD and hence this also seems to happen in a phased manner; (iii) the process of refolding seems to be reversible to the unfolding process but the protein does not regain all its structure on refolding; (iv) however, CD results reveal almost 100% recovery of the secondary structure lost during SDS induced unfolding. We have conclusively proved that there is a marginal structural gain of the native protein at low surfactant concentration and β-CD also induces a marginal structural loss to the native protein.

Exploring the self-assembly of a short aromatic Aβ(16-24) peptide.

The use of self-assembling peptides as scaffolds for creating biomaterials has prompted the scientific community to carry out studies on short peptides as model systems. Short peptides help in dissecting contributions from different interactions, unlike large peptides, where multiple interactions make it difficult to dissect the contributions of individual interactions. This opens avenues for fine tuning peptides to carry out a wide range of physical or chemical properties.

Toggling Between Blue- and Red-Emitting Fluorescent Silver Nanoclusters.

A very efficient protocol for synthesizing highly fluorescent, protein-templated silver nanoclusters (Ag/NCs) has been discussed. Two types of Ag/NCs (Ag9/HSA and Ag14/HSA), although showing significant differences in their photophysical properties, can be interconverted at will, which makes this study unique. The Ag/HSA NCs have been quantified by several spectroscopic techniques, and they find tremendous applications as photoluminescent markers.

Deciphering the role of pH in the binding of ciprofloxacin hydrochloride to bovine serum albumin.

The effect of the added fluoroquinolone, Ciprofloxacin Hydrochloride (CpH), on structural properties of Bovine Serum Albumin (BSA) was investigated by Circular Dichroism (CD), steady-state, time-resolved and Dynamic Light Scattering (DLS) spectroscopic approaches. The intrinsic fluorescence of the Tryptophan (Trp) amino acid residue in the globular protein BSA was made use of and the effect of pH at two different temperatures was thoroughly investigated. CD results indicate that CpH induces some structural changes in BSA and this has been well-supported by steady-state, lifetime and DLS data.

Protein unfolding and subsequent refolding: a spectroscopic investigation.

The mechanism by which the protein Bovine Serum Albumin (BSA) undergoes unfolding induced by Guanidine Hydrochloride (GdHCl) and then the subsequent refolding brought in by many-fold dilution was studied by steady-state fluorescence, anisotropy, time resolved measurements and Circular Dichroism (CD) spectroscopy. CD data reveal that the protein attains a degree of extra rigidity at low concentrations of the denaturant, GdHCl, and this observation was correlated with other techniques used in this present work. The unfolding and refolding of BSA appear to proceed through intermediates and both the processes are sequential in nature.

Spectroscopic determination of Critical Micelle Concentration in aqueous and non-aqueous media using a non-invasive method.

In this present study, we report on new methodology for determining the Critical Micelle Concentration (CMC) of a neutral surfactant Triton X-100 (TX-100) both in aqueous and non-aqueous media based on a non-invasive approach. The presence of the phenyl moiety of TX-100 was made use of as an intrinsic fluorophore and steady-state and time-resolved spectroscopy has been used to characterize the micellar systems. There are reports that external fluorophores may bring about some structural changes in the systems and the perturbations caused by these fluorophores in micellar systems may affect the shape and size of the micelles.

Exploring the mechanism of fluorescence quenching in proteins induced by tetracycline.

The binding of the antibiotic tetracycline hydrochloride (TC) to three proteins was investigated by steady-state, time-resolved, and circular dichroism spectroscopy. The tryptophan (Trp) amino acid residues were used as an intrinsic fluorophore to decipher the structure-function relationship. As monitored by CD spectroscopy, the addition of TC causes the protein to alter some of its helical content although such changes are only marginal.

Spectroscopic probing of the microenvironment in a protein-surfactant assembly.

The effect of the anionic surfactant sodium dodecyl sulfate (SDS) on the protein human serum albumin (HSA) was studied using steady-state spectroscopy, time-resolved measurements, and circular dichroism spectroscopy. The binding of SDS to the domain IIA of HSA, housing the single tryptophan amino acid residue (Trp214), was monitored, and it was found that this addition of the surfactant takes place in a sequential manner depending upon the concentration of the added surfactant. Both fluorescence intensity and lifetimes of HSA decreased with the increasing concentration of SDS, and the surfactant molecules serve the role of a quencher for the fluorescence of Trp214.