Established methods for characterizing proteins typically require physical or chemical modification steps or cannot be used to examine individual molecules in solution. Ionic current measurements through electrolyte-filled nanopores can characterize single native proteins in an aqueous environment, but currently offer only limited capabilities. Here we show that the zeptolitre sensing volume of bilayer-coated solid-state nanopores can be used to determine the approximate shape, volume, charge, rotational diffusion coefficient and dipole moment of individual proteins. To do this, we developed a theory for the quantitative understanding of modulations in ionic current that arise from the rotational dynamics of single proteins as they move through the electric field inside the nanopore. The approach allows us to measure the five parameters simultaneously, and we show that they can be used to identify, characterize and quantify proteins and protein complexes with potential implications for structural biology, proteomics, biomarker detection and routine protein analysis.

Real-time shape approximation and fingerprinting of single proteins using a nanopore.

The synthesis of a luminescent quantum cluster (QC) of gold with a quantum yield of approximately 4 % is reported. It was synthesized in gram quantities by the core etching of mercaptosuccinic acid protected gold nanoparticles by bovine serum albumin (BSA), abbreviated as Au(QC)@BSA. The cluster was characterized and a core of Au(38) was assigned tentatively from mass spectrometric analysis. Luminescence of the QC is exploited as a "turn-off" sensor for Cu(2+) ions and a "turn-on" sensor for glutathione detection. Metal-enhanced luminescence (MEL) of this QC in the presence of silver nanoparticles is demonstrated and a ninefold maximum enhancement is seen. This is the first report of the observation of MEL from QCs. Folic acid conjugated Au(QC)@BSA was found to be internalized to a significant extent by oral carcinoma KB cells through folic acid mediated endocytosis. The inherent luminescence of the internalized Au(QC)@BSA was used in cell imaging.

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Luminescent Quantum Clusters of Gold in Bulk by Albumin-Induced Core Etching of Nanoparticles: Metal Ion Sensing, Metal-Enhanced Luminescence, and Biolabeling

Fouling of reverse osmosis membrane by protein (BSA): Effects of pH, calcium, magnesium, ionic strength and temperature

The development of functional copper nanoclusters (Cu NCs) is becoming increasingly widespread in consumer technologies due to their applications in cellular imaging and catalysis. Herein, we report a simple protein-directed synthesis of stable, water-soluble and fluorescent Cu NCs, using BSA as the stabilising agent. Meanwhile, in this study, hydrazine hydrate (N2H4·2H2O) was used as the reducing agent. N2H4·2H2O was a mild reducing agent suggesting that all processes could be operated at room temperature. The as-prepared Cu NCs showed red fluorescence with a peaking center at 620 nm (quantum yield 4.1%). The fluorescence of the as-prepared BSA–Cu NCs was responsive to pH in that the intensity of fluorescence increased rapidly by decreasing the pH from 12 to 6. Besides, with an arresting set of features including water-dispersibility, red fluorescence, good biocompatibility, surface-bioactivity and small size, the resultant BSA–Cu NCs could be used as probes for cellular imaging and catalysis. In this study, CAL-27 cells and the reaction of oxidation of styrene are used as models to achieve fluorescence imaging and elucidate the catalytic activity of the as-prepared BSA–Cu NCs.

Protein-directed synthesis of pH-responsive red fluorescent copper nanoclusters and their applications in cellular imaging and catalysis

The adsorption of bovine serum albumin (BSA) in a planar poly(acrylic acid) (PAA) brush layer has been studied by fixed-angle optical reflectometry. The influence of polymer length, grafting density, and salt concentration is studied as a function of pH. The results are compared with predictions of an analytical polyelectrolyte brush model, which incorporates charge regulation and excluded volume interactions. A maximum in adsorption is found near the point of zero charge (pzc) of the protein. At the maximum, BSA accumulates in a PAA brush to at least 30 vol %. Substantial adsorption continues above the pzc, that is, in the pH range where a net negatively charged protein adsorbs into a negatively charged brush layer, up to a critical pH value. This critical pH value decreases with increasing ionic strength. The adsorbed amount increases strongly with both increasing PAA chain length and increasing grafting density. Experimental data compare well with the analytical model without having to include a nonhom...

Adsorption of the protein bovine serum albumin in a planar poly(acrylic acid) brush layer as measured by optical reflectometry

http://www.lps.ens.fr/~urbach/publications/ElKadiBiophysJ06.pdf

I. Nwigwe

Papers

Unfolding and Refolding of Bovine Serum Albumin at Acid pH: Ultrasound and Structural Studies