Recently a semiempirical method has been proposed by Aqvist et al. to calculate absolute and relative binding free energies. In this method, the absolute binding free energy of a ligand is estimated as deltaGbind = alpha + beta, where Vel(bound) and Vvdw(bound) are the electrostatic and van der Waals interaction energies between the ligand and t...

The extensive use of avidin and streptavidin in life sciences originates from the extraordinary tight biotin-binding affinity of these tetrameric proteins. Numerous studies have been performed to modify the biotin-binding affinity of (strept)avidin to improve the existing applications. Even so, (strept)avidin greatly favours its natural ligand, biotin. Here we engineered the biotin-binding pock...

The effect of avidin-biotin interactions in several detection systems for the non-radioactive in situ hybridization (ISH) technique was studied in a model system using a transitional cell carcinoma line and a biotinylated DNA probe. We performed fluorescence ISH to unravel the individual steps in a sensitive and frequently used amplification method which makes use of the alternating cytochemica...

This article reports on the development of a simple and cost-effective bioassay for the detection of biotin in urine and serum, based on the very selective binding of avidin and biotin. Avidin was allowed to react without isolating it from egg white. Egg white was treated with the dye HABA, which binds to avidin. Upon subsequent treatment with biotin, HABA is released due to the high affinity o...

We have examined the effect of crosslinking IgE-receptor complexes with variable receptor-receptor distances on the transmembrane signaling that leads to degranulation of rat basophilic leukemia cells. Linear polymers of the biotin-binding protein avidin were generated with bis biotin-l,12-diamidododecane, and a dinitrophenyl-biotin conjugate was bound at each end of the polymers to form a seri...

Chemomechanical functionalization of chemical surfaces is a field of chemistry and physics which has attracted a great deal of attention over the last decade. The atomic force microscope has played an instrumental role in the advancement of nanotechnology using such methods. In my research we investigated the effectiveness of depositing a stable Avidin protein monolayer on a silicon oxide surfa...

Co-immunoprecipitation of antigens from cell or tissue extracts is used in a wide variety of experimental systems to corroborate the existence of authentic protein-protein interactions in vivo. Typically, stable protein-protein associations are detected by immunoprecipitation with antibodies specific for one protein, followed by immunoblot analysis with antibodies against potential interacting ...

Fibril formation by mutational variants of human lysozyme is associated with a fatal form of hereditary non-neuropathic systemic amyloidosis. Defining the mechanistic details of lysozyme aggregation is of crucial importance for understanding the origin and progression of this disease and related misfolding conditions. In this study, we show that a biotin moiety can be introduced site-specifical...

The binding of aniohic dyes by serum albumin has been studied by many workers (reviewed by Foster, 1960). The evidence suggests several cationic sites situated in hydrophobic regions of the molecule. For example, the bound dye shows spectral changes that are in accord with its location in a non-polar environment. Since it was suggested (Green, 1963b) that the biotin-binding sites of avidin were...