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Introduction of histidine residues into avidin subunit interfaces allows pH‐dependent regulation of quaternary structure and biotin binding
Author(s) -
Nordlund Henri R,
Hytönen Vesa P,
Laitinen Olli H,
Uotila Sanna T.H,
Niskanen Einari A,
Savolainen Janne,
Porkka Eevaleena,
Kulomaa Markku S
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)01302-4
Subject(s) - avidin , histidine , protein quaternary structure , protein subunit , biotin , chemistry , biochemistry , biosensor , ligand (biochemistry) , binding site , biophysics , amino acid , biology , receptor , gene
In order to turn the subunit association and biotin binding of avidin into pH‐sensitive phenomena, we have replaced individually three amino acid residues in avidin (Met96, Val115 and Ile117) with histidines in the 1–3 interface, and in combination with a histidine conversion in the 1–2 interface (Trp110). The single replacements Met96His and Val115His in the 1–3 interface were found to have a clear effect on the quaternary structure of avidin, since subunit associations of these mutants became pH‐dependent. The histidine replacement in the 1–2 interface affected the biotin‐binding properties of the mutants, in particular reversibility of binding and protein–ligand complex formation were pH‐sensitive, as measured by IAsys biosensor and fluorescence correlation spectroscopy, respectively. The possibility of regulating the quaternary structure and function of avidin in a controlled and predictable manner, due to introduced interface histidines, will expand even further the range and versatility of the avidin–biotin technology.