Chemically synthesized 58-mer LysM domain binds lipochitin oligosaccharide

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 -