TY - JOUR
T1 - Chemically synthesized 58-mer LysM domain binds lipochitin oligosaccharide
AU - Sørensen, Kasper Kildegaard
AU - Simonsen, Jens Bæk
AU - Maolanon, Nicolai Nareth
AU - Stougaard, Jens
AU - Jensen, Knud Jørgen
PY - 2014/9/22
Y1 - 2014/9/22
N2 - Recognition of carbohydrates by proteins is a ubiquitous biochemical process. In legume-rhizobium symbiosis, lipochitin oligosaccharides, also referred to as nodulation (nod) factors, function as primary rhizobial signal molecules to trigger root nodule development. Perception of these signal molecules is receptor mediated, and nod factor receptor 5 (NFR5) from the model legume Lotus japonicus is predicted to contain three LysM domain binding sites. Here we studied the interactions between nod factor and each of the three NFR5 LysM domains, which were chemically synthesized. LysM domain variants (up to 58 amino acids) designed to optimize solubility were chemically assembled by solid-phase peptide synthesis (SPPS) with microwave heating. Their interaction with nod factors and chitin oligosaccharides was studied by isothermal titration calorimetry and circular dichroism (CD) spectroscopy. LysM2 showed a change in folding upon nod factor binding, thus providing direct evidence that the LysM domain of NFR5 recognizes lipochitin oligosaccharides. These results clearly show that the L. japonicus LysM2 domain binds to the nod factor from Mesorhizobium loti, thereby causing a conformational change in the LysM2 domain. The preferential affinity for nod factors over chitin oligosaccharides was demonstrated by a newly developed glycan microarray. Besides the biological implications, our approach shows that carbohydrate binding to a small protein domain can be detected by CD spectroscopy. A nod is as good as sugar: The L. japonicus LysM2 domain binds to the nod factor from M. loti, thereby causing a conformational change in the LysM2 domain. A glycan microarray was used to demonstrate preferential binding over oligosaccharides, and CD spectroscopy detected conformational changes.
AB - Recognition of carbohydrates by proteins is a ubiquitous biochemical process. In legume-rhizobium symbiosis, lipochitin oligosaccharides, also referred to as nodulation (nod) factors, function as primary rhizobial signal molecules to trigger root nodule development. Perception of these signal molecules is receptor mediated, and nod factor receptor 5 (NFR5) from the model legume Lotus japonicus is predicted to contain three LysM domain binding sites. Here we studied the interactions between nod factor and each of the three NFR5 LysM domains, which were chemically synthesized. LysM domain variants (up to 58 amino acids) designed to optimize solubility were chemically assembled by solid-phase peptide synthesis (SPPS) with microwave heating. Their interaction with nod factors and chitin oligosaccharides was studied by isothermal titration calorimetry and circular dichroism (CD) spectroscopy. LysM2 showed a change in folding upon nod factor binding, thus providing direct evidence that the LysM domain of NFR5 recognizes lipochitin oligosaccharides. These results clearly show that the L. japonicus LysM2 domain binds to the nod factor from Mesorhizobium loti, thereby causing a conformational change in the LysM2 domain. The preferential affinity for nod factors over chitin oligosaccharides was demonstrated by a newly developed glycan microarray. Besides the biological implications, our approach shows that carbohydrate binding to a small protein domain can be detected by CD spectroscopy. A nod is as good as sugar: The L. japonicus LysM2 domain binds to the nod factor from M. loti, thereby causing a conformational change in the LysM2 domain. A glycan microarray was used to demonstrate preferential binding over oligosaccharides, and CD spectroscopy detected conformational changes.
KW - Glycan microarray
KW - Glycobiology
KW - Microwave chemistry
KW - Plant-microbe interactions
KW - Solid-phase synthesis
U2 - 10.1002/cbic.201402125
DO - 10.1002/cbic.201402125
M3 - Journal article
AN - SCOPUS:84908155205
SN - 1439-4227
VL - 15
SP - 2097
EP - 2105
JO - ChemBioChem
JF - ChemBioChem
IS - 14
ER -