Engineering of charge, biotin-binding and oligomerization of avidin : new tools for avidin-biotin technology

Abstract

Avidin is a tetrameric glycoprotein found in chicken egg white. Due to its remarkably high affinity for biotin, avidin has widely been used in biological and biotechnical applications in life sciences. Avidin is also a highly stable protein, maintaining its functional tetrameric structure at high temperatures, extremes of pH, and in the presence of strong denaturants. In the present study we wanted to find out whether different properties of avidin can be modified with protein engineering using rational design. We used the 3D-structure of avidin together with sequence information from various avidin-like proteins to plan these changes. Besides shedding more light on the protein chemical bases for the high-affinity binding, stability or the physicochemical characteristics of avidin, our aim was also to create new tools for the avidin-biotin technology.