A1 Refereed original research article in a scientific journal

Chicken avidin-related protein 4/5 shows superior thermal stability when compared with avidin while retaining high affinity to biotin




AuthorsHytonen VP, Nyholm TKM, Pentikainen OT, Vaarno J, Porkka EJ, Nordlund HR, Johnson MS, Slotte JP, Laitinen OH, Kulomaa MS

PublisherAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Publication year2004

JournalJournal of Biological Chemistry

Journal name in sourceJOURNAL OF BIOLOGICAL CHEMISTRY

Journal acronymJ BIOL CHEM

Volume279

Issue10

First page 9337

Last page9343

Number of pages7

ISSN0021-9258

DOIhttps://doi.org/10.1074/jbc.M310989200(external)


Abstract
The protein chicken avidin is a commonly used tool in various applications. The avidin gene belongs to a gene family that also includes seven other members known as the avidin-related genes (AVR). We report here on the extremely high thermal stability and functional characteristics of avidin-related protein AVR4/5, a member of the avidin protein family. The thermal stability characteristics of AVR4/5 were examined using a differential scanning calorimeter, microparticle analysis, and a microplate assay. Its biotin-binding properties were studied using an isothermal calorimeter and IAsys optical biosensor. According to these analyses, in the absence of biotin AVR4/5 is clearly more stable (T-m = 107.4 +/- 0.3 degreesC) than avidin (T-m = 83.5 +/- 0.1 degreesC) or bacterial streptavidin (T-m = 75.5 degreesC). AVR4/5 also exhibits a high affinity for biotin (K-d approximate to 3.6 x 10(-14) M) comparable to that of avidin and streptavidin (K-d approximate to 10(-15) M). Molecular modeling and site-directed mutagenesis were used to study the molecular details behind the observed high thermostability. The results indicate that AVR4/5 and its mutants have high potential as new improved tools for applications where exceptionally high stability and tight biotin binding are needed.



Last updated on 2024-26-11 at 10:39