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Acute monocytic leukemia is a type of myeloid leukemia that develops in monocytes. The current clinical therapies for leukemia are unsatisfactory due to their side effects and nonspecificity toward target cells. Some lectins display antitumor activity and may specifically recognize cancer cells by binding to carbohydrate structures on their surface. Therefore, this study evaluated the response of the human monocytic leukemia cell lines THP-1 to the Olneya tesota PF2 lectin. The induction of apoptosis and reactive oxygen species production in PF2-treated cells was evaluated by flow cytometry, and the lectin-THP-1 cell interaction and mitochondrial membrane potential were evaluated by confocal fluorescence microscopy. PF2 genotoxicity was evaluated by DNA fragmentation analysis via gel electrophoresis. The results showed that PF2 binds to THP-1 cells, triggers apoptosis and DNA degradation, changes the mitochondrial membrane potential, and increases reactive oxygen species levels in PF2-treated THP-1 cells. These results suggest the potential use of PF2 for developing alternative anticancer treatments with enhanced specificity., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

Prohibitin (PHB) is a highly conserved eukaryotic protein complex involved in multiple cellular processes. In insects, PHB has been identified as a potential target protein to insecticidal molecules acting as a receptor of PF2 insecticidal lectin in the midgut of Zabrotes subfasciatus larvae (bean pest) and Cry protein of Bacillus thuringiensis in Leptinotarsa decemlineata (Colorado potato beetle). This study aimed to characterize the structural features of Z. subfasciatus prohibitin (ZsPHB) by homology modeling and evaluate its expression and tissue localization at different stages of larval development both at the transcript and protein levels. The samples were collected from eggs and larvae of different developmental stages. The immunodetection of ZsPHB was done with anti-PHB1 and confirmed by LC-MS/MS analysis. Gene expression analysis of ZsPHB1 and ZsPHB2 was performed by RT-qPCR, and immunohistochemistry with FITC-labeled anti-PHB1. Results showed that ZsPHBs exhibit distinctive characteristics of the SPFH protein superfamily. The transcript levels suggest a coordinated expression of ZsPHB1 and ZsPHB2 genes, while ZsPHB1 was detected in soluble protein extracts depending on the stage of development. Histological examination showed ZsPHB1 is present in all larval tissues, with an intense fluorescence signal observed at the gut. These results suggest a physiologically important role of PHB during Z. subfasciatus development and show its regulation occurs at the transcriptional and post-transcriptional levels. This is the first characterization of PHB in Z. subfasciatus., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: The O. tesota lectin PF2 is a tetrameric protein with subunits of 33 kDa that recognizes only complex carbohydrates, resistant to proteolytic enzymes and has insecticidal activity against Phaseolus beans pest., Objective: To explore PF2 lectin features at different protein structural levels and to evaluate the effect of temperature and pH on its functionality and conformational stability., Methods: PF2 lectin was purified by affinity chromatography. Its primary structure was resolved by mass spectrometry and analyzed by bioinformatic tools, including its tertiary structure homology modeling. The effect of temperature and pH on its conformational traits and stability was addressed by dynamic light scattering, circular dichroism, and intrinsic fluorescence. The hemagglutinating activity was evaluated using a suspension of peripheral blood erythrocytes., Results: The proposed PF2 folding comprises a high content of beta sheets. At pH 7 and 25°C, the hydrodynamic diameter (Dh) was found to be 12.3 nm which corresponds to the oligomeric native state of PF2 lectin. Dh increased under the other evaluated pH and temperature conditions, suggesting protein aggregation. At basic pH, PF2 exhibited low conformational stability. The native PF2 (pH 7) retained its full hemagglutinating activity up to 45°C and exhibited one transition state with a melting temperature of 76.8°C., Conclusion: PF2 showed distinctive characteristics found in legume lectins. The pH influences the functionality and conformational stability of the protein. PF2 lectin displayed a relatively narrow thermostability to the loss of secondary structure and hemagglutinating activity., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The available genomic and proteomic information of non-model organisms is often underrepresented in public databases hindering their study at molecular, cellular, and physiological levels. Information on Zabrotes subfasciatus (Mexican bean weevil) is poorly represented in databases, yet it is a major pest of common beans. We report the transcriptome of Z. subfasciatus larvae; transcripts were sequenced using an Illumina RNA-Seq technology and assembled de novo identifying 29,029 unigenes with an average size of 1168 bp and an N50 value of 2196 bp. About 15,124 unigenes (52%) were functionally annotated and categorized. Further analysis revealed 30 unigene sequences encoding putative targets of the insecticidal PF2 lectin. The complete deduced amino acid sequences of eight selected proteins potentially related to insecticidal mechanism of Palo Fierro 2 (PF2) were used for predicting probable N-glycosylation sites and analyzing phylogenetic relationships with insect sequences. This work provides a dramatic increase in the genetic resources available for Coleopterans and set the basis for developing future studies on biological aspects and potential control strategies for Z. subfasciatus .

Lectins are a diverse class of proteins distributed extensively in nature. Among these proteins; legume lectins display a variety of interesting features including antimicrobial; insecticidal and antitumor activities. Because lectins recognize and bind to specific glycoconjugates present on the surface of cells and intracellular structures; they can serve as potential target molecules for developing practical applications in the fields of food; agriculture; health and pharmaceutical research. This review presents the current knowledge of the main structural characteristics of legume lectins and the relationship of structure to the exhibited specificities; provides an overview of their particular antimicrobial; insecticidal and antitumor biological activities and describes possible applications based on the pattern of recognized glyco-targets., Competing Interests: The authors declare no conflict of interest.

Zabrotes subfasciatus (Boheman) is the main pest of common beans ( Phaselous vulgaris ). Wild legume seeds from Olneya tesota contain a lectin, PF2, that shows insecticidal activities against this insect. The binding of PF2 to midgut glycoproteins of 20-day-old larvae was evaluated using PF2 affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the proteins retained on the gel revealed several putative glycoproteins, ranging in mass from 17 to 97 kDa. Subsequent protein digestion and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) provided amino acid fragments that identified an α-tubulin, cytochrome c oxidase subunit I, an odorant receptor, and a lysozyme from available insect sequence databases. The potential of these proteins to serve as part of the mechanisms involved in the insecticidal activity of PF2 to Z. subfasciatus is discussed.

Zabrotes subfasciatus (Boheman) is the main pest of common beans (Phaselous vulgaris). Some wild legume seeds may contain lectins with insecticidal activities against this insect. The larval developments of Z. subfasciatus on seeds of Olneya tesota (a desert wild legume) and on artificial seeds containing purified PF2 lectin were evaluated. PF2 susceptibility to proteolysis was assessed by incubation with midgut extract at different times. PF2 binding to midgut glycoconjugates was assessed by histochemistry. A reduced level of oviposition and a lack of emergence of adult beetles were observed in O. tesota seeds (compared to common beans), and in artificial seeds containing PF2 at 0.5 and 1%. PF2 was resistant to larval midgut proteolysis for 24 h, while PHA-E (lectin control) was fully digested after 4 h. Histochemistry analysis of midguts incubated with PF2 showed recognition for microvillae and possibly with peritrophic gel. On the other hand, PHA-E exhibited no interaction with larval midgut glycoproteins. Proteolysis resistance and glycan recognition could in part explain why PF2 is toxic to Z. subfasciatus while PHA is not.