Crystallization and preliminary X-ray diffraction analysis of a new chitin-binding protein from<i>Parkia platycephala</i>seeds

Cavada, Benildo S.; Castellón, Rolando E. R.; Vasconcelos, Georg G.; Rocha, B.A.M.; Bezerra, G.A.; Debray, Henri; Delatorre, P.; Nagano, Celso Shiniti; Toyama, Marcos H.; Pinto, Vicente de Paulo Teixeira; Moreno, Frederico Bruno Mendes Batista; Canduri, Fernanda; Azevedo, Walter Filgueira de

doi:10.1107/s1744309105024462

Cited by 4 publications

(4 citation statements)

References 19 publications

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“…PPL2 was crystallized by the hanging drop vapor diffusion method at 20 °C as described [28]. The crystals belong to the P2 1 2 1 2 1 space group with one monomer in the asymmetric unit.…”

Section: Methodsmentioning

confidence: 99%

“…fungi) secreting pathogenesis‐related enzymes, such as xylanases and chitinases, which degrade the pathogen's cell wall [25–27]. In a previous article we have reported the presence of an endochitinase in Parkia platycephala seeds [28]. Now, we have determined its complete amino acid sequence by a combination of Edman degradation and cDNA cloning, and report its biochemical characterization and the determination of its crystal structure.…”

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confidence: 99%

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cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N‐acetylglucosamine‐binding hemagglutinin from Parkia platycephala seeds

et al. 2006

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Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP-HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407 ± 15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N-acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed b(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-b-d-glucopyranose units in chitin. The full-length amino acid sequence of Parkia platycephala lectin 2, determined by N-terminal sequencing and cDNA cloning, and its three-dimensional structure, established by X-ray crystallography at 1.75 Å resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (ba) 8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182.Abbreviations CTAB, cetyl triethylammonium bromide; GlcNac, N-acetyl-D-glucosamine; GSP, gene-specific forward primer; HPAEC-PAD, high-pH anion exchange chromatography with pulsed amperometric detection; PE, pyridylethylated; PPL1, Parkia platycephala lectin 1; PPL2, Parkia platycephala lectin 2; PTC, phenylisothiocyanate; PTH, phenylthiohydantoin.

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“…PPL2 was crystallized by the hanging drop vapor diffusion method at 20 °C as described [28]. The crystals belong to the P2 1 2 1 2 1 space group with one monomer in the asymmetric unit.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N‐acetylglucosamine‐binding hemagglutinin from Parkia platycephala seeds

et al. 2006

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“…Other cPLA2 enzymes include diacylglycerol (DAG), which is involved in protein kinase C (PKC) and phospholipase C (PLC) activation. This activation is responsible for the production of inositol triphosphate (IP 3 ) and DAG [ 18 , 19 , 20 , 21 ]. Notwithstanding, PKC and PLC are also key elements in pathway signaling mediated by protease-activated receptors (PARs).…”

Section: Introductionmentioning

confidence: 99%

Edema Induced by a Crotalus durissus terrificus Venom Serine Protease (Cdtsp 2) Involves the PAR Pathway and PKC and PLC Activation

Costa

Belchor

Rodrigues

et al. 2018

IJMS

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Snake venom serine proteases (SVSPs) represent an essential group of enzymatic toxins involved in several pathophysiological effects on blood homeostasis. Some findings suggest the involvement of this class of enzymatic toxins in inflammation. In this paper, we purified and isolated a new gyroxin isoform from the Crotalus durissus terrificus (Cdt) venom, designated as Cdtsp 2, which showed significant proinflammatory effects in a murine model. In addition, we performed several studies to elucidate the main pathway underlying the edematogenic effect induced by Cdtsp 2. Enzymatic assays and structural analysis (primary structure analysis and three-dimensional modeling) were closely performed with pharmacological assays. The determination of edematogenic activity was performed using Cdtsp 2 isolated from snake venom, and was applied to mice treated with protein kinase C (PKC) inhibitor, phospholipase C (PLC) inhibitor, dexamethasone (Dexa), antagonists for protease-activated receptors (PARs), or saline (negative control). Additionally, we measured the levels of cyclooxygenase 2 (COX-2), malondialdehyde (MDA), and prostaglandin E2 (PGE2). Cdtsp 2 is characterized by an approximate molecular mass of 27 kDa, an isoelectric point (pI) of 4.5, and significant fibrinolytic activity, as well as the ability to hydrolyze Nα-benzoyl-l-arginine 4-nitroanilide (BAPNA). Its primary and three-dimensional structures revealed Cdtsp 2 as a typical snake venom serine protease that induces significant edema via the metabolism of arachidonic acid (AA), involving PARs, PKC, PLC, and COX-2 receptors, as well as inducing a significant increase in MDA levels. Our results showed that Cdtsp 2 is a serine protease with significant enzymatic activity, and it may be involved in the degradation of PAR1 and PAR2, which activate PLC and PKC to mobilize AA, while increasing oxidative stress. In this article, we provide a new perspective for the role of SVSPs beyond their effects on blood homeostasis.

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“…Class I has an N-terminal domain, which is similar to a domain present in hevein, a protein from rubber latex , whereas class II PR-4 proteins lack this domain. Examples of such chitin-binding proteins are chitinases from bean , wheat , and tobacco , chitin-binding lectins from wheat , barley , rice , breadfruit , Pakia platycephala , chitin-binding proteins and peptides from Amaranthus retroflexus and Amaranthus caudatus , , sugar beet , Pharbitis nil L. , Eucommia ulmoides , and Ginkgo biloba . The size of chitin-binding proteins varies from 3.1 up to 20 kDa; they often have a basic pI and are highly resistant to extreme pH and protease treatment .…”

Section: Introductionmentioning

confidence: 99%

Isolation and Biochemical Characterization of an Antifungal Peptide from Amaranthus hypochondriacus Seeds

Rivillas‐Acevedo

Soriano-Garcı́a

2007

J. Agric. Food Chem.

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An antifungal peptide, Ay-AMP, was isolated from Amaranthus hypochondriacus seeds by acidic extraction and then purified by reverse-phase high-pressure liquid chromatography. The molecular mass of this peptide, as determined by mass spectrometry, is 3184 Da. The peptide belongs to the superfamily of chitin-binding proteins, containing a single cysteine/glycine-rich chitin-binding domain, and it was found that Ay-AMP degrades chitin. Ay-AMP inhibits the growth, at very low doses, of different pathogenic fungi, such as Candida albicans, Trichoderma sp., Fusarium solani, Penicillium chrysogenum, Geotrichum candidum, Aspergillus candidus, Aspergillus schraceus, and Alternaria alternata. Ay-AMP is very resistant to the effect of proteases and heating; however, it showed an antagonistic effect with CaCl2 and KCl.

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Crystallization and preliminary X-ray diffraction analysis of a new chitin-binding protein fromParkia platycephalaseeds

Cited by 4 publications

References 19 publications

cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N‐acetylglucosamine‐binding hemagglutinin from Parkia platycephala seeds

cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N‐acetylglucosamine‐binding hemagglutinin from Parkia platycephala seeds

Edema Induced by a Crotalus durissus terrificus Venom Serine Protease (Cdtsp 2) Involves the PAR Pathway and PKC and PLC Activation

Isolation and Biochemical Characterization of an Antifungal Peptide from Amaranthus hypochondriacus Seeds

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