Backbone dynamics of the antifungal Psd1 pea defensin and its correlation with membrane interaction by NMR spectroscopy.
Plant defensins are cysteine-rich cationic peptides, components of the innate immune system. The antifungal sensitivity of certain exemplars was correlated to the level of complex glycosphingolipids in the membrane of fungi strains. Psd1 is a 46 amino acid residue defensin isolated from pea seeds which exhibit antifungal activity. Its structure is characterized by the so-called cysteine-stabilized alpha/beta motif linked by three loops as determined by two-dimensional NMR. In the present work we explored the measurement of heteronuclear Nuclear Overhauser Effects, R1 and R2 (15)N relaxation ratios, and chemical shift to probe the backbone dynamics of Psd1 and its interaction with membrane mimetic systems with phosphatidylcholine (PC) or dodecylphosphocholine (DPC) with glucosylceramide (CMH) isolated from Fusarium solani. The calculated R2 values predicted a slow motion around the highly conserved among Gly12 residue and also in the region of the Turn3 His36-Trp38. The results showed that Psd1 interacts with vesicles of PC or PC:CMH in slightly different forms. The interaction was monitored by chemical shift perturbation and relaxation properties. Using this approach we could map the loops as the binding site of Psd1 with the membrane. The major binding epitope showed conformation exchange properties in the mus-ms timescale supporting the conformation selection as the binding mechanism. Moreover, the peptide corresponding to part of Loop1 (pepLoop1: Gly12 to Ser19) is also able to interact with DPC micelles acquiring a stable structure and in the presence of DPC:CMH the peptide changes to an extended conformation, exhibiting NOE mainly with the carbohydrate and ceramide parts of CMH.
de Medeiros
Luciano Neves
LN
Instituto de Biofísica Carlos Chagas Filho, Programa de Biologia Molecular e Estrutural, Universidade Federal do Rio de Janeiro, Brazil.
Angeli
Renata
R
Sarzedas
Carolina G
CG
Barreto-Bergter
Eliana
E
Valente
Ana Paula
AP
Kurtenbach
Eleonora
E
Almeida
Fabio C L
FC
D020816:Amino Acid Motifs / Q000502:physiology; D023082:Defensins / Q000737:chemistry* / Q000378:metabolism; D005670:Fusarium / Q000737:chemistry* / Q000378:metabolism; D008567:Membranes, Artificial; D008823:Micelles; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D018532:Pisum sativum / Q000737:chemistry* / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
D023082:Defensins; D008567:Membranes, Artificial; D008823:Micelles; D010743:Phospholipids; D010940:Plant Proteins; C410643:defensin 1, Pisum sativum