6UD1

Spectroscopic and structural characterization of a genetically encoded direct sensor for protein-ligand interactions

6UD1 の概要

• エントリーDOI: 10.2210/pdb6ud1/pdb
• 分子名称: Streptavidin, CHLORIDE ION (3 entities in total)
• 機能のキーワード: unnatural amino acid, non-canonical amino acid, fluorescence, biosensor, small molecule biosensor, ligand detection, fluorescent protein
• 由来する生物種: Streptomyces avidinii
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 29156.32

構造登録者

Mills, J.H.,Gleason, P.R.,Simmons, C.R.,Henderson, J.N.,Kartchner, B.K. (登録日: 2019-09-18, 公開日: 2021-02-10, 最終更新日: 2024-10-30)

主引用文献

Gleason, P.R.,Kolbaba-Kartchner, B.,Henderson, J.N.,Stahl, E.P.,Simmons, C.R.,Mills, J.H.
Structural Origins of Altered Spectroscopic Properties upon Ligand Binding in Proteins Containing a Fluorescent Noncanonical Amino Acid.
Biochemistry, 60:2577-2585, 2021

PubMed Abstract: Fluorescent noncanonical amino acids (fNCAAs) could serve as starting points for the rational design of protein-based fluorescent sensors of biological activity. However, efforts toward this goal are likely hampered by a lack of atomic-level characterization of fNCAAs within proteins. Here, we describe the spectroscopic and structural characterization of five streptavidin mutants that contain the fNCAA l-(7-hydroxycoumarin-4-yl)ethylglycine (7-HCAA) at sites proximal to the binding site of its substrate, biotin. Many of the mutants exhibited altered fluorescence spectra in response to biotin binding, which included both increases and decreases in fluorescence intensity as well as red- or blue-shifted emission maxima. Structural data were also obtained for three of the five mutants. The crystal structures shed light on interactions between 7-HCAA and functional groups, contributed either by the protein or by the substrate, that may be responsible for the observed changes in the 7-HCAA spectra. These data could be used in future studies aimed at the rational design of fluorescent, protein-based sensors of small molecule binding or dissociation.

PubMed: 34415744
DOI: 10.1021/acs.biochem.1c00291

実験手法

X-RAY DIFFRACTION (1.55 Å)