14 août 19251925/08/14 (A47,N16469).Appartient à l’ensemble documentaire : ChArdenn

Bessolicina E.

Ermakova E.

Khairutdinov B.

Kovaleva V.

Nesmelova I.

Tarasova N.

Toporkova Y.

Yusypovych Y.

Zuev Y.

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

© 2017 Elsevier B.V.Plants have developed a complex defense response system against pests and pathogens. Defensins, produced by plants as part of their innate immune response, form the family of small, basic, cysteine-rich proteins with activity primarily directed against fungal pathogens. In addition, plant defensins can show antibacterial activity and protease and insect amylase inhibitory activities. However, in gymnosperms, only antifungal activity of defensins has been described thus far. Here, we report antibacterial and insect α-amylase inhibition activities for defensin PsDef1 from P. sylvestris, the first defensin from gymnosperms with a broad range of biological activities described. We also report the solution NMR structure of PsDef1 and its dynamics properties assessed by a combination of experimental NMR and computational techniques. Collectively, our data provide an insight into structure, dynamics, and functional properties of PsDef1 that could be common between defensins from this taxonomic group

Kazan Federal University Digital Repository

NMR structure, conformational dynamics, and biological activity of PsDef1 defensin from Pinus sylvestris

Contents lists available at ScienceDirectBBA - Proteins and Proteomicsjournal homepage: www.elsevier.com/locate/bbapapNMR structure, conformational dynamics, and biological activity of PsDef1defensin from Pinus sylvestrisBulat I. Khairutdinova,b,1, Elena A. Ermakovaa,1, Yuri M. Yusypovychc, Elena K. Bessolicinaa,Nadezhda B. Tarasovaa, Yana Y. Toporkovaa, Valentina Kovalevac, Yuriy F. Zueva,b,d,Irina V. Nesmelovae,f,⁎a Kazan Institute of Biochemistry and Biophysics, Lobachevsky St. 2/31, Kazan 420111, Russiab Kazan Federal University, Kremlyovskaya Str. 18, Kazan 420008, Russiac Ukrainian National Forestry University, Gen. Chyprunku St. 103, Lviv 79057, Ukrained Kazan State Power Engineering University, Krasnoselskaya st. 51, Kazan 420066, Russiae Department of Physics and Optical Science, University of North Carolina, 9201 University City Blvd., Charlotte, NC 28223, USAf Center for Biomedical Engineering and Science, University of North Carolina, 9201 University City Blvd., Charlotte, NC 28223, USAA R T I C L E I N F OKeywords:Scotts pine defensinPlant defensinNMR structureConformational dynamicsAntibacterial activityAmylase inhibitionA B S T R A C TPlants have developed a complex defense response system against pests and pathogens. Defensins, produced byplants as part of their innate immune response, form the family of small, basic, cysteine-rich proteins withactivity primarily directed against fungal pathogens. In addition, plant defensins can show antibacterial activityand protease and insect amylase inhibitory activities. However, in gymnosperms, only antifungal activity ofdefensins has been described thus far. Here, we report antibacterial and insect α-amylase inhibition activities fordefensin PsDef1 from P. sylvestris, the ﬁrst defensin from gymnosperms with a broad range of biological activitiesdescribed. We also report the solution NMR structure of PsDef1 and its dynamics properties assessed by acombination of experimental NMR and computational techniques. Collectively, our data provide an insight intostructure, dynamics, and functional properties of PsDef1 that could be common between defensins from thistaxonomic group.1. IntroductionDefense mechanisms of plants against pests and pathogens includethe production of antimicrobial peptides (AMPs). The group of plantAMPs that resemble structural and functional features of previouslycharacterized antimicrobial peptides found in insects and mammals, arecalled defensins [1]. Most plant defensins show activity against fungi[2–4], but they can also act as inhibitors of protein synthesis [5,6], α-amylases [7–9], proteases [10,11], or ion channels [12]. Some plantdefensins possess the activity against Gram-positive and Gram-negativebacteria [13,14].Plant defensins are small, cysteine-rich, cationic peptides of ap-proximately 45–54 amino acid residues [1,15,16]. The amino acidsequence identity between diﬀerent plant defensins can be< 35%[2,17] and only a small number of residues is conserved. Most plantdefensins have six or eight conserved cysteine residues that form threeand four disulﬁde bonds, respectively, although a few plant defensinswith ten cysteine residues and ﬁve disulphide bonds have beenidentiﬁed [16]. In addition to conserved cysteine residues, a glycine,a serine, and an aromatic residue (F/W/H/Y) that is always followed byanother glycine are conserved. Conserved disulﬁde bonds are thoughtto deﬁne the physico-chemical properties of defensins, such as anextreme resistance to high temperatures and acidic environments[18–21]. The stability of plant defensins makes them attractive asbiotechnological tools against phytopathogenic fungi, currently con-trolled only by chemicals [2].Despite the large variability of amino acid sequence, all plantdefensins with reported three-dimensional structures show a similarthree-dimensional fold as monomers, consisting of a triple-strandedantiparallel β-sheet and one α-helix tethered to the β-sheet by disulﬁdebonds, i.e., a cysteine-stabilized αβ, or CSαβ fold [2]. Two antiparallelβ-strands β2 and β3 joined by a loop (loop L3) form a γ-core motifGXCX3–9C (X being any amino acid and G being a conserved glycineresidue). The γ-core motif has a net positive charge and is thought to behttp://dx.doi.org/10.1016/j.bbapap.2017.05.012Received 6 March 2017; Received in revised form 14 May 2017; Accepted 16 May 2017⁎ Corresponding author at: Grigg Hall 306, Department of Physics and Optical Sciences, University of North Carolina, 9201 University City Blvd., Charlotte, NC 28223, USA.1 Authors contributed equally to this work.E-mail address: Irina.Nesmelova@uncc.edu (I.V. Nesmelova).BBA - Proteins and Proteomics 1865 (2017) 1085–1094Available online 17 May 20171570-9639/ © 2017 Elsevier B.V. All rights reserved.MARK

Gallica, Bibliotheque Numerique