Calcium-dependent hydrolysis of supported planar lipids was triggered by honey bee venom phospholipase A<sub>2</sub>with the right orientation at the interface

Kai, Siqi; Li, Xu; Han, Xiaofeng; Lü, Xiaolin

doi:10.1039/c7cp06344j

Cited by 11 publications

(25 citation statements)

References 28 publications

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“…Here in particular, the secondary signal was chosen to be a poison for the receiver GUV so that a predator–prey relationship was established between the two GUV populations. The predator GUVs were loaded with membrane impermeable Ca 2+ ions, which upon transfer through the α-HL pores to the prey GUV activate the phospholipase A 2 (PLA 2 ) , and lead to a fluorescent signal coming from the calcium sensitive dye, Rhod2. Finally, the calcium activated PLA 2 cleaved the fatty acid tail of the phospholipid in the prey GUVs and caused to their collapse.…”

Section: Resultsmentioning

confidence: 99%

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Chakraborty

Wegner

2021

ACS Nano

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Cells commonly communicate with each other through diffusible molecules but nonchemical communication remains elusive. While bioluminescent organisms communicate through light to find prey or attract mates, it is still under debate if signaling through light is possible at the cellular level. Here, we demonstrate that cell to cell signaling through light is possible in artificial cell communities derived from biomimetic vesicles. In our design, artificial sender cells produce an intracellular light signal, which triggers the adhesion to receiver cells. Unlike soluble molecules, the light signal propagates fast, independent of diffusion and without the need for a transporter across membranes. To obtain a predator–prey relationship, the luminescence predator cells is loaded with a secondary diffusible poison, which is transferred to the prey cell upon adhesion and leads to its lysis. This design provides a blueprint for light based intercellular communication, which can be used for programing artificial and natural cell communities.

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Section: Resultsmentioning

confidence: 99%

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Chakraborty

Wegner

2021

ACS Nano

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“…With the help of molecular dynamics in combination with mass spectrometry, it was shown that the lipid membrane works as an allosteric activator for sPLA2 [8]; binding to the membrane leads to conformational changes and activates the enzyme [9]. Using a combination of confocal microscopy and IR spectrometry, it was shown that calcium ions can be involved in the binding of sPLA2 to the membrane surface [10]; calcium ions can orient protein molecules on the membrane surface and activate the enzyme. Moreover, fluorescence microscopy and atomic force microscopy studies revealed that changes in the membrane structure caused by sPLA2 lead to an increase in the amount of adsorbed protein, that is, to the formation of new lipid binding sites for sPLA2 [11].…”

Section: Short Communicationmentioning

confidence: 99%

Estimation of the Phospholipase A2 Selectivity on POPC/POPG Membranes Using the Interaction Map

Алексеева

Volynsky

Болдырев

2021

Biochem. Moscow Suppl. Ser. A

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The regulation of the activity and selectivity of phospholipase A2 (PLA2), which is capable of cleaving fatty acid in the second position (sn-2) of the phospholipid, is carried out through the membrane-binding and catalytic sites of the enzyme. For hydrolytic activity, PLA2 must first bind to the phospholipid membrane, and the binding efficiency depends on the composition of the membrane. The membrane-binding site of PLA2 is formed by several tens of amino acids and its composition differs from enzyme to enzyme; hydrophobic and positively charged amino acids play a key role in the interaction. In this work, we investigated the interaction of PLA2 from bee venom with phospholipid bilayers of palmitoyl oleoylphosphatidylcholine (POPC) containing different amounts of palmitoyloleoylphosphatidylglycerol (POPG). On the basis of the measurements of the protein intrinsic fluorescence and the anisotropy of the fluorescence of the lipid probe we propose the construction of lipid–protein interaction maps, which reflect both the efficiency of protein binding and changes in the structure of the membrane. These changes cause alterations in the fluorescence anisotropy of the label, which in turn is a measure of the mobility of the lipid environment of the fluorescent probe. Analysis of interaction maps showed that there is a relationship between lipid mobility and enzyme binding efficiency: the optimum interaction of PLA2 with membranes from a POPC/POPG mixture lies in the region of the highest lipid mobility, and not in the region of the highest negative charge. This dependence complements the existing understanding of the process of recognition of the membrane surface by the enzyme and the selection of lipids by the enzyme already bound to the membrane. The proposed mapping method can be extended to other membrane-active proteins.

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“…Measuring the direct effect of phospholipases on lipid bilayers using such impedance measurements has also been described, but the techniques require a time-consuming lipid bilayer deposition methodology or include the use of toxic solvents such as chloroform . Likewise, other techniques such as the use of a quartz crystal microbalance, sum-frequency generation vibrational spectroscopy measurements, and atomic force microscopy have been used to study the mechanistic properties of PLA isoforms on lipid bilayers, − but they also rely on lengthy substrate deposition methodologies. To the best of our knowledge, most phospholipase biosensor designs are also incapable of rapidly distinguishing between phospholipase isoforms such as phospholipase A 1 (PLA 1 ) and phospholipase A 2 (PLA 2 ).…”

Section: Introductionmentioning

confidence: 99%

Label-Free, Real-Time Phospholipase-A Isoform Assay

García

Deplazes

Aili

et al. 2020

ACS Biomater. Sci. Eng.

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Phospholipase-A (PLA) enzymes catalyze the hydrolysis of ester bonds in select glycerophospholipids. Sensors for rapidly measuring the PLA activity in biological samples have relevance in the study of venom compositions and in medical diagnostics for the diagnosis of diseases such as acute pancreatitis. Current PLA sensor technologies are often restricted by the time it takes to prepare an assay, the necessity of using fluorescent labels, or the fact they might require strict pH control of the buffer vehicles used. Here we present a tethered bilayer lipid membrane (tBLM) impedance sensor array for the rapid and real-time detection of PLA, which includes the ability to selectively detect phospholipase-A 2 (PLA 2 ) from phospholipase-A 1 (PLA 1 ) isoforms. Comparing the activity of PLA 1 and PLA 2 in an array of tBLMs composed of ether phospholipids, ester phospholipids or ether-ester phospholipids allows for the rapid and reliable distinction between the isoforms, as measured using swept-frequency electrical impedance spectroscopy. After testing the assay using pure enzymes, we demonstrate the capacity of the sensor to identify specific PLA 2 -type, calcium-dependent activity from the venom of the South American bullet ant, Paraponera clavata, at a concentration of 1 μg/mL. The specificity of the phospholipase activity was corroborated using matrixassisted laser-desorption/ionization time-of-flight mass spectrometry. As further validation, we tested the activities of a PLA 1 isoform in the presence of different buffers commonly used in biology and biochemistry experiments. Sensitivity testing shows that PLA 1 can be detected at an activity as low as 0.06 U/mL. The rapid and reliable detection of phospholipases presented in this study has potential applications in the study of animal venoms as well as in lipase bioreactors and point-of-care devices.

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Calcium-dependent hydrolysis of supported planar lipids was triggered by honey bee venom phospholipase A₂with the right orientation at the interface

Cited by 11 publications

References 28 publications

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Estimation of the Phospholipase A2 Selectivity on POPC/POPG Membranes Using the Interaction Map

Label-Free, Real-Time Phospholipase-A Isoform Assay

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Calcium-dependent hydrolysis of supported planar lipids was triggered by honey bee venom phospholipase A2with the right orientation at the interface

Cited by 11 publications

References 28 publications

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Cell to Cell Signaling through Light in Artificial Cell Communities: Glowing Predator Lures Prey

Estimation of the Phospholipase A2 Selectivity on POPC/POPG Membranes Using the Interaction Map

Label-Free, Real-Time Phospholipase-A Isoform Assay

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Product

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Calcium-dependent hydrolysis of supported planar lipids was triggered by honey bee venom phospholipase A₂with the right orientation at the interface