Next Article in Journal
Insights into the Effect of the G245S Single Point Mutation on the Structure of p53 and the Binding of the Protein to DNA
Next Article in Special Issue
Solution NMR Spectroscopy in Target-Based Drug Discovery
Previous Article in Journal
Synthesis of 5′-GalNAc-Conjugated Oligonucleotides: A Comparison of Solid and Solution-Phase Conjugation Strategies
Previous Article in Special Issue
A Unique and Simple Approach to Improve Sensitivity in 15N-NMR Relaxation Measurements for NH3+ Groups: Application to a Protein-DNA Complex
Article Menu
Issue 8 (August) cover image

Export Article

Open AccessFeature PaperArticle
Molecules 2017, 22(8), 1357; doi:10.3390/molecules22081357

Lactose Binding Induces Opposing Dynamics Changes in Human Galectins Revealed by NMR-Based Hydrogen–Deuterium Exchange

1
Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan
2
Department of Chemistry, National Taiwan University, Taipei 106, Taiwan
3
Institute of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan
*
Author to whom correspondence should be addressed.
Received: 26 July 2017 / Revised: 8 August 2017 / Accepted: 10 August 2017 / Published: 16 August 2017
(This article belongs to the Special Issue Recent Advances in Biomolecular NMR Spectroscopy)
View Full-Text   |   Download PDF [3688 KB, uploaded 16 August 2017]   |  

Abstract

Galectins are β-galactoside-binding proteins implicated in a myriad of biological functions. Despite their highly conserved carbohydrate binding motifs with essentially identical structures, their affinities for lactose, a common galectin inhibitor, vary significantly. Here, we aimed to examine the molecular basis of differential lactose affinities amongst galectins using solution-based techniques. Consistent dissociation constants of lactose binding were derived from nuclear magnetic resonance (NMR) spectroscopy, intrinsic tryptophan fluorescence, isothermal titration calorimetry and bio-layer interferometry for human galectin-1 (hGal1), galectin-7 (hGal7), and the N-terminal and C-terminal domains of galectin-8 (hGal8NTD and hGal8CTD, respectively). Furthermore, the dissociation rates of lactose binding were extracted from NMR lineshape analyses. Structural mapping of chemical shift perturbations revealed long-range perturbations upon lactose binding for hGal1 and hGal8NTD. We further demonstrated using the NMR-based hydrogen–deuterium exchange (HDX) that lactose binding increases the exchange rates of residues located on the opposite side of the ligand-binding pocket for hGal1 and hGal8NTD, indicative of allostery. Additionally, lactose binding induces significant stabilisation of hGal8CTD across the entire domain. Our results suggested that lactose binding reduced the internal dynamics of hGal8CTD on a very slow timescale (minutes and slower) at the expense of reduced binding affinity due to the unfavourable loss of conformational entropy. View Full-Text
Keywords: galectins; protein dynamics; NMR spectroscopy; ligand binding galectins; protein dynamics; NMR spectroscopy; ligand binding
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Chien, C.-T.H.; Ho, M.-R.; Lin, C.-H.; Hsu, S.-T.D. Lactose Binding Induces Opposing Dynamics Changes in Human Galectins Revealed by NMR-Based Hydrogen–Deuterium Exchange. Molecules 2017, 22, 1357.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]

Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top