Structural Characterization of the Avidin Interactions with Fluorescent Pyrene-Conjugates: 1-Biotinylpyrene and 1-Desthiobiotinylpyrene†Reportar como inadecuado


Structural Characterization of the Avidin Interactions with Fluorescent Pyrene-Conjugates: 1-Biotinylpyrene and 1-Desthiobiotinylpyrene†


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1

Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, 90-924 Łódź, Stefanowskiego 4-10, Poland

2

Department of Organic Chemistry, Faculty of Chemistry, University of Lodz, 91-403 Łódź, Tamka 12, Poland



This article is dedicated to Alexander Wlodawer on the occasion of his 70th birthday.





*

Author to whom correspondence should be addressed.



Academic Editor: Derek J. McPhee

Abstract Avidin is a tetrameric protein that belongs to the calycin superfamily. It has been studied mainly because of its extraordinary affinity to biotin, which led to a wide range of applications based on the avidin-biotin system. In the present study, we report the first crystal structures of avidin in a complex with two novel fluorescent pyrene derivatives: 1-biotinylpyrene B9P and 1-desthiobiotinylpyrene D9P. The crystal structures were solved by molecular replacement using the coordinates of avidin molecule as a starting model and the final models of avidin-B9P and avidin-D9P were refined to resolutions of 2.0 Å and 2.1 Å, respectively. Our data reveal changes in loop conformation as well as in overall fold and quaternary arrangement of the avidin upon the binding of these fluorescent probes. Moreover, the crystal structures allowed analysis of the details of the interactions between the protein and the pyrene derivatives. Structural description of the complexes will contribute to the design of conjugates for expanding the capabilities of avidin–biotin technology. View Full-Text

Keywords: crystal structure; avidin; pyrene; biotin; 1-biotinylpyrene; 1-desthiobiotinylpyrene; protein crystallography; protein-ligand interaction crystal structure; avidin; pyrene; biotin; 1-biotinylpyrene; 1-desthiobiotinylpyrene; protein crystallography; protein-ligand interaction





Autor: Paweł Strzelczyk 1, Damian Plażuk 2, Janusz Zakrzewski 2 and Grzegorz Bujacz 1,*

Fuente: http://mdpi.com/



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