Dataset Information

Aggregation and Molecular Properties of ?-Glucosidase Isoform II in Chayote (Sechium edule).

ABSTRACT: The presence of isoforms of ?-glucosidase has been reported in some grasses such as sorghum, rice and maize. This work aims to extract and characterize isoform II in ?-glucosidase from S. edule. A crude extract was prepared without buffer solution and adjusted to pH 4.6. Contaminating proteins were precipitated at 4 °C for 24 h. The supernatant was purified by chromatography on carboxymethyl cellulose (CMC) column, molecular exclusion on Sephacryl S-200HR, and exchange anionic on QFF column. Electrophoretic analyzes revealed a purified enzyme with aggregating molecular complex on SDS-PAGE, Native-PAGE, and AU-PAGE. Twelve peptides fragments were identified by nano liquid chromatography-tandem mass spectrometry (nano LC-ESI-MS/MS), which presented as 61% identical to Cucurbita moschata ?-glucosidase and 55.74% identical to ?-glucosidase from Cucumis sativus, another Cucurbitaceous member. The relative masses which contained 39% hydrophobic amino acids ranged from 982.49 to 2,781.26. The enzyme showed a specificity to ?-d-glucose with a Km of 4.59 mM, a Vmax value of 104.3 ?M?min-1 and a kcat of 10,087 ?M?min-1 using p-nitrophenyl-?-D-glucopyranoside. The presence of molecular aggregates can be attributed to non-polar amino acids. This property is not mediated by a ?-glucosidase aggregating factor (BGAF) as in grasses (maize and sorghum). The role of these aggregates is discussed.

SUBMITTER: Cruz Rodriguez A

PROVIDER: S-EPMC7180739 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Publications

Aggregation and Molecular Properties of β-Glucosidase Isoform II in Chayote (<i>Sechium edule</i>).

Cruz Rodríguez Alberto A Sánchez Esperanza Fabiola Anaid FA Pérez-Campos Eduardo E Hernández-Huerta María Teresa MT Pérez-Campos Mayoral Laura L Matias-Cervantes Carlos Alberto CA Martínez Barras Alexis A Mayoral-Andrade Gabriel G Santos Pineda Luis Ángel LÁ Díaz Barrita Aymara Judith AJ Zenteno Edgar E Romero Díaz Carlos C Martínez Cruz Ruth R Pérez-Campos Mayoral Eduardo E Bernabé Pérez Edith Alhelí EA Pérez Santiago Alma Dolores AD Pina-Canseco María Del Socorro MDS Martínez Cruz Margarito M

Molecules (Basel, Switzerland) 20200408 7

The presence of isoforms of β-glucosidase has been reported in some grasses such as sorghum, rice and maize. This work aims to extract and characterize isoform II in β-glucosidase from <i>S. edule</i>. A crude extract was prepared without buffer solution and adjusted to pH 4.6. Contaminating proteins were precipitated at 4 °C for 24 h. The supernatant was purified by chromatography on carboxymethyl cellulose (CMC) column, molecular exclusion on Sephacryl S-200HR, and exchange anionic on QFF colu ...[more]

PMID: 32276317

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Project description:A comparison between the axon terminals of octopaminergic efferent dorsal or ventral unpaired median neurons in either desert locusts (Schistocerca gregaria) or fruit flies (Drosophila melanogaster) across skeletal muscles reveals many similarities. In both species the octopaminergic axon forms beaded fibers where the boutons or varicosities form type II terminals in contrast to the neuromuscular junction (NMJ) or type I terminals. These type II terminals are immunopositive for both tyramine and octopamine and, in contrast to the type I terminals, which possess clear synaptic vesicles, only contain dense core vesicles. These dense core vesicles contain octopamine as shown by immunogold methods. With respect to the cytomatrix and active zone peptides the type II terminals exhibit active zone-like accumulations of the scaffold protein Bruchpilot (BRP) only sparsely in contrast to the many accumulations of BRP identifying active zones of NMJ type I terminals. In the fruit fly larva marked dynamic changes of octopaminergic fibers have been reported after short starvation which not only affects the formation of new branches ("synaptopods") but also affects the type I terminals or NMJs via octopamine-signaling (Koon et al., 2011). Our starvation experiments of Drosophila-larvae revealed a time-dependency of the formation of additional branches. Whereas after 2 h of starvation we find a decrease in "synaptopods", the increase is significant after 6 h of starvation. In addition, we provide evidence that the release of octopamine from dendritic and/or axonal type II terminals uses a similar synaptic machinery to glutamate release from type I terminals of excitatory motor neurons. Indeed, blocking this canonical synaptic release machinery via RNAi induced downregulation of BRP in neurons with type II terminals leads to flight performance deficits similar to those observed for octopamine mutants or flies lacking this class of neurons (Brembs et al., 2007).

Dataset Information

Aggregation and Molecular Properties of ?-Glucosidase Isoform II in Chayote (Sechium edule).

Publications

Aggregation and Molecular Properties of β-Glucosidase Isoform II in Chayote (<i>Sechium edule</i>).

Similar Datasets

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