Structure of bradavidin-C-terminal residues act as intrinsic ligands.

PloS One
Jenni LeppiniemiTomi T Airenne

Abstract

Bradavidin is a homotetrameric biotin-binding protein from Bradyrhizobium japonicum, a nitrogen fixing and root nodule-forming symbiotic bacterium of the soybean. Wild-type (wt) bradavidin has 138 amino acid residues, whereas the C-terminally truncated core-bradavidin has only 118 residues. We have solved the X-ray structure of wt bradavidin and found that the C-terminal amino acids of each subunit were uniquely bound to the biotin-binding pocket of an adjacent subunit. The biotin-binding pocket occupying peptide (SEKLSNTK) was named "Brad-tag" and it serves as an intrinsic stabilizing ligand in wt bradavidin. The binding of Brad-tag to core-bradavidin was analysed by isothermal titration calorimetry and a binding affinity of ∼25 µM was measured. In order to study the potential of Brad-tag, a green fluorescent protein tagged with Brad-tag was prepared and successfully concentrated from a bacterial cell lysate using core-bradavidin-functionalized Sepharose resin.

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Citations

May 11, 2013·Protein Science : a Publication of the Protein Society·Jenni LeppiniemiVesa P Hytönen
Jul 2, 2015·Journal of Structural Biology·Orly AvrahamOded Livnah
Apr 21, 2017·PloS One·Nitin AgrawalTomi T Airenne
Mar 24, 2018·The Journal of General Physiology·Keith K Khoo, Stephan A Pless

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Methods Mentioned

BETA
X-ray
isothermal titration calorimetry
biolayer
biosensor
biosensors
biolayer interferometry
peptide array
biosensing
PCR

Software Mentioned

Mosflm
Data Analysis
PyMOL
Coot
Data Acquisition
iMosflm
Scala
wARP
Phenix
PyMOL Molecular Graphics System

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