Determination of the three-dimensional solution structure of Raphanus sativus Antifungal Protein 1 by 1 H NMR 1 1Edited by P. E. Wright

Fant, Franky; Vranken, Wim; Broekaert, Willem F.; Borremans, Frans

doi:10.1006/jmbi.1998.1767

Cited by 151 publications

(156 citation statements)

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“…Due to the undesirable side-effect of synthetic agro-chemicals many biologists have been intensively studying biological control of plant fungal pathogens [1][2][3][4][5][6][7][8][9] . Antimicrobial proteins such as radish antifungal protein in radish seeds, RIP (ribosome inactivating protein) and zeamatin in many plant seeds have been reported in the previous studies 1,[10][11][12][13][14] . Several types of AMPs have been isolated, and their structures have been fully or partially characterized.…”

Section: Introductionmentioning

confidence: 99%

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

Hwang¹

2009

Korean Journal of Environmental Agriculture

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In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

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Section: Introductionmentioning

confidence: 99%

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

Hwang¹

2009

Korean Journal of Environmental Agriculture

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“…Genes of plant defensins are expressed in different organs, e.g. seeds [7,8], leaves [9], roots [10], flowers [11,12] and symbiotic tubercles [13]. There is both constitutive and inducible expression of genes, encoding plant defensins [11,14].…”

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confidence: 99%

“…There is both constitutive and inducible expression of genes, encoding plant defensins [11,14]. The main function of defensins is to inhibit fungal disease [8,11,15], but for some peptides there are antibacterial activity [16,17], inhibition of trypsin [18], and participation in the formation of resistance to heavy metals [19], cold stress [20,21], drought [22], salinity [23,24] and in developmental processes [25].…”

mentioning

confidence: 99%

“…Using nuclear-magnetic resonance spectroscopy, it was shown that in plant defensins the basis of the molecule is formed by a three-layer element consisting of one -spiral connected with cysteine bridges to -bands (Fig. 1) [7,8,27, 28]. The lines connecting the cysteine residues are disulfide bridges.…”

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confidence: 99%

“…AlfAFP from the seeds of Medicago sativa is antifungal peptide and provides resistance to fungal pathogen Verticillium dahlia in transgenic potato plants [11,31]. Defensins from Raphanus sativus (RsAFP) are accumulated as the seeds ripen and are released after the integrity of seed membranes is destroyed to create a microenvironment that suppresses the growth of fungi, with a decrement in the elongation of hyphae and their enhanced branching [8,32]. MsDef1 from M. sativa seeds significantly inhibits the growth of F. graminearum in vitro [33].…”

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PLANT DEFENSINS: BIOLOGICAL FUNCTION, MECHANISMS OF ACTION AND METHODS OF ANALYSIS (review)

Kulaeva¹,

Kliukova²,

Повыдыш³

et al. 2017

S-h. biol.

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A b s t r a c tOne of the actual problems of modern agriculture is crop loss due to various biotic and abiotic factors. In plants there is a multicomponent protection system, including the formation of protective barriers, activation the reaction of hypersensitivity and synthesis of antimicrobial peptides, which are low molecular weight compounds showing broad spectrum activity against fungi, bacteria and viruses. This group consists of several groups of peptides, including defensins, which are one of the most common classes of antimicrobial peptides and are detectable in all living organisms. Defensins are small (45-54 amino acids), cysteine-rich peptides involved in a different protective responses (B.P. Thomma et al., 2002). Genes which are coding plant defensins are expressed in different organs of plants, where their products are necessary for biotic and abiotic stresses. Thus, these peptides are extremely important in terms of getting crop lines that are resistant to pathogens and abiotic stresses. Defensins are characterized by a strong sequence variability that seems to correlate with a variety of mechanisms of action of these peptides that can induce pathogen's cell death by penetrating into a cell or by being on its surface (T.M. Shafee et al., 2016). Most of the plant defensins are characterized by the antifungal activity. Some defensins have antibacterial activity, which may be combined with activity against fungi. For a small number of plant defensins their participation in the processes of resistance to heavy metals, cold stress, drought, salinity, and in the development process is indicated. Modern approaches of molecular and computational biology allow an effective search for new forms of defensins activity by studying the wild, non-model plant objects. The development of next-generation sequencing methods («Next Generation Sequencing») make possible the intensive study of the transcriptomes of such objects. However, the correct annotation of the sequences of peptides, characterized by small size and high variability, can be done by usage the special programs, such as SPADA (Small Peptide Alignment Discovery Application) (P. Zhou et al., 2013). SPADA makes multiple sequence alignment of all known paralogous genes within a gene family and builds a predictive model for the search of new members of the same family. Prediction of newly identified active defensins and identification of conserved amino acids can also be performed by computational biology methods. An approach based on a multiple sequence alignment and subsequent cluster analysis allows dividing defensins into groups with similar functional activity (N.L. van der Weerden et al., 2013). Thus, the combination of modern methods of molecular and computational biology allows carrying out the study of this group of protective peptides with high efficiency.

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The three-dimensional solution structure ofAesculus hippocastanum antimicrobial protein 1 determined by1H nuclear magnetic resonance

Fant¹,

Vranken²,

Borremans³

1999

Proteins

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Aesculus hippocastanum antimicrobial protein 1 (Ah-AMP1) is a plant defensin isolated from horse chestnuts. The plant defensins have been divided in several subfamilies according to their amino acid sequence homology. Ah-AMP1, belonging to subfamily A2, inhibits growth of a broad range of fungi. So far, a three-dimensional structure has been determined only for members of subfamilies A3 and B2. In order to understand activity and specificity of these plant defensins, the structure of a protein belonging to subfamily A2 is needed. We report the three-dimensional solution structure of Ah-AMP1 as determined from twodimensional 1 H nuclear magnetic resonance data. The structure features all the characteristics of the ''cysteine-stabilized ␣␤-motif.'' A comparison of the structure, the electrostatic potential surface and regions important for interaction with the fungal receptor, is made with Rs-AFP1 (plant defensin of subfamily A3). Thus, residues important for activity and specificity have been assigned. Proteins 1999;37:388-403.

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Determination of the three-dimensional solution structure of Raphanus sativus Antifungal Protein 1 by 1 H NMR 1 1Edited by P. E. Wright

Cited by 151 publications

References 59 publications

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

PLANT DEFENSINS: BIOLOGICAL FUNCTION, MECHANISMS OF ACTION AND METHODS OF ANALYSIS (review)

The three-dimensional solution structure ofAesculus hippocastanum antimicrobial protein 1 determined by1H nuclear magnetic resonance

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