Crystal structure of vespid phospholipase A1 reveals insights into the mechanism for cause of membrane dysfunction

Hou, Ming-Hon; Chuang, Chia-Yi; Ko, Tzu‐Ping; Hu, Nien-Jen; Chou, Chia Cheng; Shih, Yan-Ping; Ho, C.L.; Wang, Andrew H.‐J.

doi:10.1016/j.ibmb.2015.11.002

Cited by 20 publications

(29 citation statements)

References 30 publications

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“…Phospholipase A1 is a major allergen widely distribute in social Hymenoptera venoms (Santos et al, 2011;Hou et al, 2016). Vespid phospholipases A1 (vPLA1s) are known to hydrolyze ester bonds of 1,2-diacyl-3-sn glycerophospholipids at the sn-1 position, converting these substrates into their corresponding lyso compounds and releasing fatty acids (Santos et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…Vespid phospholipases A1 (vPLA1s) are known to hydrolyze ester bonds of 1,2-diacyl-3-sn glycerophospholipids at the sn-1 position, converting these substrates into their corresponding lyso compounds and releasing fatty acids (Santos et al, 2007). During the envenoming process, vPLA1s are able to disrupt the phospholipid packing of biological membranes, causing severe hemolysis and leading to cardiac dysfunction and death in animals (Hou et al, 2016). The allergen showed direct hemolytic action against washed red blood cells (Santos et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Molecular cloning, expression and IgE-immunoreactivity of phospholipase A1, a major allergen from Polybia paulista (Hymenoptera: Vespidae) venom

Pérez-Riverol

Pereira

Lasa

et al. 2016

Toxicon

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Polybia paulista (Hymenoptera: Vespidae) is a clinically relevant social wasp that frequently causes stinging accidents in southeast Brazil. To date, diagnosis and specific immunotherapy (SIT) of allergy are based on the use of crude venom extracts. Production of recombinant forms of major allergens from P. paulista venom will improve diagnosis and SIT of allergic patients by reducing the incidence of cross-reactivity and non-specific sensitization. Here, we describe the molecular cloning, heterologous expression, purification and IgE-mediated immunodetection of phospholipase A1 (Poly p 1), a major allergen from P. paulista venom. The cDNA of Poly p 1 was extracted from venom glands and then cloned, and further expression of the recombinant allergen (rPoly p 1) was achieved in Escherichia coli BL21 (DE3) cells. Purification of rPoly p 1 was performed using immobilized Ni metal affinity chromatography. Also, a single-step chromatographic method allowed the purification of native Poly p 1 (nPoly p 1) from the wasp's venom glands. We used western blotting to evaluate IgE-reactivity of the sera from 10 P. paulista venom-allergic patients to rPoly p 1 and nPoly p 1. High levels of insoluble rPoly p 1 were obtained during heterologous expression. After solubilization of inclusion bodies and purification of the recombinant protein, a unique band of ∼34 kDa was detected in SDS-PAGE analysis. Allergen-specific IgE (sIgE) from allergic patients' sera recognized rPoly p 1, nPoly p 1 and crude venom extract to a similar extent. Our results showed that rPoly p 1 could be used for development of component-resolved diagnosis (CRD) and molecular-defined SIT of P. paulista venom allergy.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular cloning, expression and IgE-immunoreactivity of phospholipase A1, a major allergen from Polybia paulista (Hymenoptera: Vespidae) venom

Pérez-Riverol

Pereira

Lasa

et al. 2016

Toxicon

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“…It thus is reasonable to assume that high-level intracellular expression of foreign PLA1 can disturb the balanced asymmetry and deteriorate the integrity of the cell membrane. This assumption is also strengthened by the fact that exogenous PLA1s secreted from pathogenic bacteria, and those found in the venom of bees and snakes, cause destabilization and hemolysis of nearly all kinds of cells [ 22 , 23 ].…”

Section: Resultsmentioning

confidence: 99%

Cell-free synthesis of functional phospholipase A1 from Serratia sp.

Lim

Park

Jang

et al. 2016

Biotechnol Biofuels

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BackgroundPhospholipase A1 is an enzyme that hydrolyzes phospholipids at the sn-1 position. It has potential applications across diverse fields including food, pharmaceutical, and biofuel industries. Although there has been increasing interest in the use of phospholipase A1 for degumming of plant oils during biodiesel production, production of recombinant phospholipase A1 has been hampered by low efficiency of gene expression and its toxicity to the host cell.ResultsWhile expression of phospholipase A1 in Escherichia coli resulted in extremely low productivity associated with inhibition of transformed cell growth, drastically higher production of functional phospholipase A1 was achieved in a cell-free protein synthesis system where enzyme expression is decoupled from cell physiology. Compared with expression in E. coli, cell-free synthesis resulted in an over 1000-fold higher titer of functional phospholipase A1. Cell-free produced phospholipase A1 was also used for successfully degumming crude plant oil.ConclusionsWe demonstrate successful production of Serratia sp. phospholipase A1 in a cell-free protein synthesis system. Including the phospholipase A1 investigated in this study, many industrial enzymes can interfere with the regular physiology of cells, making cellular production of them problematic. With the experimental results presented herewith, we believe that cell-free protein synthesis will provide a viable option for rapid production of important industrial biocatalysts.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0563-5) contains supplementary material, which is available to authorized users.

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“…Venom PLA2s activity could produce edematogenic, hypotensive, cytotoxic and anticoagulant effects in the victims [84]. Meanwhile, some wasp PLA1s have showed a potent hemolytic activity [85] although their major consequence during envenoming is the induction of allergic reactions [35]. The role of these P. paulista venom toxins along with those involved in inflammation (mastoparans Polybia MPI, II, III, Polybia CP, 2, 3) and allergic reactions (Poly p 1, 2 and 5) mentioned in the proposed mechanism of envenoming will be addressed later in this review.…”

Section: Exploring P Paulista Venom -Omics Approachesmentioning

confidence: 99%

Wasp venomic: Unravelling the toxins arsenal of Polybia paulista venom and its potential pharmaceutical applications

Pérez-Riverol

Santos-Pinto

Lasa

et al. 2017

Journal of Proteomics

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Polybia paulista (Hymenoptera: Vespidae) is a neotropical social wasp from southeast Brazil. As most social Hymenoptera, venom from P. paulista comprises a complex mixture of bioactive toxins ranging from low molecular weight compounds to peptides and proteins. Several efforts have been made to elucidate the molecular composition of the P. paulista venom. Data derived from proteomic, peptidomic and allergomic analyses has enhanced our understanding of the whole envenoming process caused by the insect sting. The combined use of bioinformatics, -omics- and molecular biology tools have allowed the identification, characterization, in vitro synthesis and recombinant expression of several wasp venom toxins. Some of these P. paulista - derived bioactive compounds have been evaluated for the rational design of antivenoms and the improvement of allergy specific diagnosis and immunotherapy. Molecular characterization of crude venom extract has enabled the description and isolation of novel toxins with potential biotechnological applications. Here, we review the different approaches that have been used to unravel the venom composition of P. paulista. We also describe the main groups of P. paulista - venom toxins currently identified and analyze their potential in the development of component-resolved diagnosis of allergy, and in the rational design of antivenoms and novel bioactive drugs.

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Crystal structure of vespid phospholipase A1 reveals insights into the mechanism for cause of membrane dysfunction

Cited by 20 publications

References 30 publications

Molecular cloning, expression and IgE-immunoreactivity of phospholipase A1, a major allergen from Polybia paulista (Hymenoptera: Vespidae) venom

Molecular cloning, expression and IgE-immunoreactivity of phospholipase A1, a major allergen from Polybia paulista (Hymenoptera: Vespidae) venom

Cell-free synthesis of functional phospholipase A1 from Serratia sp.

Wasp venomic: Unravelling the toxins arsenal of Polybia paulista venom and its potential pharmaceutical applications

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