Soybean ubiquitous urease with purification facilitator: An addition to the moonlighting studies toolbox

Published on Feb 1, 2017in Process Biochemistry2.883
· DOI :10.1016/j.procbio.2016.12.003
Anne H.S. Martinelli9
Estimated H-index: 9
(UFRGS: Universidade Federal do Rio Grande do Sul),
Fernanda Cortez Lopes9
Estimated H-index: 9
(PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)
+ 12 AuthorsCélia R. Carlini31
Estimated H-index: 31
(PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)
Abstract Ureases are nickel-dependent enzymes that catalyze the hydrolysis of urea to ammonia and carbon dioxide. In soybean ( Glycine max ), the embryo-specific urease (eSBU), the ubiquitous urease (uSBU), and a third isoform (SBU-III) are synthesized. Our group has previously demonstrated that eSBU, purified from seeds, has antifungal properties against phytopathogenic fungi, entomotoxicity against Dysdercus peruvianus , the ability to induce blood platelet aggregation, and these properties are independent of its enzymatic activity. Here we describe the biological properties of apo-uSBU fused to glutathione S-transferase (GST) produced in Escherichia coli . Removal of GST affected apo-uSBU stability. We performed a Response Surface Methodology to optimize GST-uSBU production to 5 mg per liter and then bioassays were carried out. The recombinant protein exhibited inhibitory effects on filamentous fungi and affected fungal secondary metabolism. Candida albicans and C. tropicalis were also susceptible to GST-uSBU and formed pseudo-hyphae. The fusion protein was toxic against Rhodnius prolixus, with the toxicity being accompanied by in vivo and in vitro hemocyte aggregation. Rabbit platelet also aggregated in the presence of GST-uSBU. Thus, uSBU displayed similar biological properties as previously described for eSBU even when fused to GST, reinforcing the proposed role of ureases in plant defense.
  • References (61)
  • Citations (2)
📖 Papers frequently viewed together
63 Citations
26 Citations
32 Citations
78% of Scinapse members use related papers. After signing in, all features are FREE.
#1Beatriz Wiebke-Strohm (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 8
#2Rodrigo Ligabue-Braun (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 11
Last. Maria Helena Bodanese-Zanettini (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 21
view all 6 authors...
Abstract In plants, ureases have been related to urea degradation, to defense against pathogenic fungi and phytophagous insects, and to the soybean− Bradyrhizobium japonicum symbiosis. Two urease isoforms have been described for soybean: the embryo-specific, encoded by Eu 1 gene, and the ubiquitous urease, encoded by Eu 4. A third urease-encoding locus exists in the completed soybean genome. The gene was designated Eu5 and the putative product of its ORF as SBU-III. Phylogenetic analysis shows t...
6 CitationsSource
#1Tiago Tezotto (USP: University of São Paulo)H-Index: 11
#2Sarah Caroline Ribeiro de Souza (State University of Campinas)H-Index: 5
Last. Joseph C. Polacco (MU: University of Missouri)H-Index: 35
view all 7 authors...
The soybean eu3-a mutant (formerly, eu3-e1) lacks all ureolytic activity. Eu3 encodes urease accessory (Ni insertion) protein, UreG. Eu3 (Glycine max v1.1 Glyma08g08970.1) is the only UreG-encoding gene in the soybean genome. Here we show that the eu3-a lesion is a 2.4 kb deletion, beginning 50 bp upstream of the transcription start, and covering 94 % of the deduced amino acid sequence of UreG, explaining the urease activity-null phenotype of eu3-a. We established near-isogenic lines (NILs), eu3...
5 CitationsSource
#1Célia R. Carlini (PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)H-Index: 31
#2Rodrigo Ligabue-Braun (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 11
Ureases are metalloenzymes that hydrolyze urea into ammonia and carbon dioxide. They were the first enzymes to be crystallized and, with them, the notion that enzymes are proteins became accepted. Novel toxic properties of ureases that are independent of their enzyme activity have been discovered in the last three decades. Since our first description of the neurotoxic properties of canatoxin, an isoform of the jack bean urease, which appeared in Toxicon in 1981, about one hundred articles have b...
39 CitationsSource
#2Célia R. CarliniH-Index: 31
2 CitationsSource
#1Marina S. Defferrari (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 9
#2R. da Silva (McMaster University)H-Index: 1
Last. Célia R. Carlini (PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)H-Index: 31
view all 4 authors...
Ureases are multifunctional proteins that display biological activities independently of their enzymatic function, such as induction of exocytosis and insecticidal effects. Rhodnius prolixus, a major vector of Chagas' disease, is a model for studies on the entomotoxicity of jack bean urease (JBU). We have previously shown that JBU induces the production of eicosanoids in isolated tissues of R. prolixus. In insects, the immune response comprises cellular and humoral reactions, and is centrally mo...
6 CitationsSource
#1Mahmoud Fahmi Elsebai (University of Oulu)H-Index: 12
#2Muhammad Saleem (IUB: Islamia University)H-Index: 37
Last. Anna Maria Pirttilä (University of Oulu)H-Index: 28
view all 8 authors...
Covering up to the end of August 2013 Phenalenones are members of a unique class of natural polyketides exhibiting diverse biological potential. This is a comprehensive review of 72 phenalenones with diverse structural features originating from fungal sources. Their bioactive potential and structure elucidation are discussed along with a review of their biosynthetic pathways and the taxonomical relationship between the fungi producing these natural products.
25 CitationsSource
#1Anne H.S. Martinelli (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 9
#2Karine Kappaun (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 3
Last. Giancarlo Pasquali (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 22
view all 13 authors...
Abstract Background Ureases are metalloenzymes involved in defense mechanisms in plants. The insecticidal activity of Canavalia ensiformis (jack bean) ureases relies partially on an internal 10 kDa peptide generated by enzymatic hydrolysis of the protein within susceptible insects. A recombinant version of this peptide, jaburetox, exhibits insecticidal, antifungal and membrane-disruptive properties. Molecular modeling of jaburetox revealed a prominent β-hairpin motif consistent with either neuro...
25 CitationsSource
#1Christos Papaneophytou (UTH: University of Thessaly)H-Index: 12
#2George Kontopidis (UTH: University of Thessaly)H-Index: 23
Abstract The supply of many valuable proteins that have potential clinical or industrial use is often limited by their low natural availability. With the modern advances in genomics, proteomics and bioinformatics, the number of proteins being produced using recombinant techniques is exponentially increasing and seems to guarantee an unlimited supply of recombinant proteins. The demand of recombinant proteins has increased as more applications in several fields become a commercial reality. Escher...
68 CitationsSource
#1Anuradha Balasubramanian (University of Madras)H-Index: 6
#2Vishnuprabu Durairajpandian (University of Madras)H-Index: 1
Last. Karthe Ponnuraj (University of Madras)H-Index: 13
view all 6 authors...
Urease is an enzyme that catalyzes the hydrolysis of urea, forming ammonia and carbon dioxide, and is found in plants, microorganisms and invertebrates. Although plant and bacterial ureases are closely related at amino acid and at the structural level, the insecticidal activity is seen only in the plant ureases. In contrast, both plant and bacterial ureases exhibit antifungal activity. These two biological properties are independent of its ureolytic activity. However, till date the mechanism(s) ...
25 CitationsSource
#1Rafael Real-GuerraH-Index: 7
Last. Célia R. CarliniH-Index: 31
view all 3 authors...
Ureases are metalloproteins responsible for the one step hydrolysis of urea into ammonia and carbamate [1], the later then rapidly and spontaneously decomposes to form carbon di‐ oxide and a second molecule of ammonia [2]. Plant ureases hold a special place in science history, participating on some important landmarks of biochemistry. For instance, it was by the analysis of Canavalia ensiformis urease crystals that the proteinaceous nature of enzymes could be demonstrated [3], rendering a Chemis...
6 CitationsSource
Cited By2
#1Karine Kappaun (PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)H-Index: 3
#2Anne H.S. Martinelli (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 9
Last. Stefano CiurliH-Index: 37
view all 11 authors...
Ureases from different biological sources display non-ureolytic properties that contribute to plant defense, in addition to their classical enzymatic urea hydrolysis. Antifungal and entomotoxic effects were demonstrated for Jaburetox, an intrinsically disordered polypeptide derived from jack bean (Canavalia ensiformis) urease. Here we describe the properties of Soyuretox, a polypeptide derived from soybean (Glycine max) ubiquitous urease. Soyuretox was fungitoxic to Candida albicans, leading to ...
2 CitationsSource
#1Karine Kappaun (PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)H-Index: 3
#2Angela Regina Piovesan (PUCRS: Pontifícia Universidade Católica do Rio Grande do Sul)H-Index: 4
Last. Rodrigo Ligabue-Braun (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 11
view all 4 authors...
Abstract Urease (urea amidohydrolase, EC is a nickel-containing enzyme produced by plants, fungi, and bacteria that catalyzes the hydrolysis of urea into ammonia and carbamate. Urease is of historical importance in Biochemistry as it was the first enzyme ever to be crystallized (1926). Finding nickel in urease’s active site (1975) was the first indication of a biological role for this metal. In this review, historical and structural features, kinetics aspects, activation of the metalloc...
8 CitationsSource
#1Angela Menegassi (UFRGS: Universidade Federal do Rio Grande do Sul)H-Index: 2
Last. Arlete Beatriz (ULBRA: Universidade Luterana do Brasil)H-Index: 7
view all 5 authors...
Ureases catalyze the hydrolysis of urea into ammonia and carbon dioxide and, thus, are involved in the metabolism and bioavailability of nitrogen. Ureases occur in plants, fungi, and bacteria. In plants, besides their enzymatic activity, ureases as proteins play a role in defense against insect and phytopathogenic fungi. Little is known about the regulation of urease in plants under stress and whether or not phytohormones may be involved. In this study, we addressed the regulation of ubiquitous ...
1 CitationsSource