In vitro refolded napin-like protein of Momordica charantia expressed in Escherichia coli displays properties of native napin

Vashishta, Aruna; Sahu, Tejram; Sharma, Anshu; Choudhary, Sunil K.; Dixit, Aparna

doi:10.1016/j.bbapap.2006.03.010

Cited by 14 publications

(8 citation statements)

References 39 publications

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“…However, it shared some extent of sequence similarity with other plant proteins, including napin also from BG (Fig. 1f) [19,20] and Ricinus communis (castor beans) 2S albumin [21], and others.…”

Section: Resultsmentioning

confidence: 95%

“…c Thirdly, fraction SP3 was pooled, dialyzed, and finally loaded on a Superdex 75 column connected to an AKTA FPLC system. Elution was carried out in 0.05 M NH 4 HCO 3 buffer, and RNase MC2 was eluted as a homogeneous peak ( [19,20]; CM Cucurbita maxima (winter squash) 2S albumin, RC Ricinus communis (castor bean) 2S albumin (PRF: 227594) [21]. Charantin, a 9.7 kDa ribosome inactivating protein from M. charantia (BG) [48] Apoptosis (2012) 17:377-387 379…”

Section: Assays Of Ribonucleolytic Activity and Base-specificitymentioning

confidence: 99%

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RNase MC2: a new Momordica charantia ribonuclease that induces apoptosis in breast cancer cells associated with activation of MAPKs and induction of caspase pathways

et al. 2011

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Ribonucleases (RNases) are ubiquitously distributed nucleases that cleave RNA into smaller pieces. They are promising drugs for different cancers based on their concrete antitumor activities in vitro and in vivo. Here we report for the first time purification and characterization of a 14-kDa RNase, designated as RNase MC2, in the seeds of bitter gourd (Momordica charantia). RNase MC2 manifested potent RNA-cleavage activity toward baker's yeast tRNA, tumor cell rRNA, and an absolute specificity for uridine. RNase MC2 demonstrated both cytostatic and cytotoxic activities against MCF-7 breast cancer cells. Treatment of MCF-7 cells with RNase MC2 caused nuclear damage (karyorrhexis, chromatin condensation, and DNA fragmentation), ultimately resulting in early/late apoptosis. Further molecular studies unveiled that RNase MC2 induced differential activation of MAPKs (p38, JNK and ERK) and Akt. On the other hand, RNase MC2 exposure activated caspase-8, caspase-9, caspase-7, increased the production of Bak and cleaved PARP, which in turn contributed to the apoptotic response. In conclusion, RNase MC2 is a potential agent which can be exploited in the worldwide fight against breast cancer.

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Section: Resultsmentioning

confidence: 95%

Section: Assays Of Ribonucleolytic Activity and Base-specificitymentioning

confidence: 99%

RNase MC2: a new Momordica charantia ribonuclease that induces apoptosis in breast cancer cells associated with activation of MAPKs and induction of caspase pathways

et al. 2011

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“…The major challenge in the expression of properly folded 2S albumins is the formation of the four disulfide bridges. Previously, recombinant 2S albumins from various species have been successfully expressed in both the yeast Pichia pastoris 35–38 and E. coli 15–17, 39–43. Three forms of hazelnut 2S albumin were cloned and expressed (not shown): the full‐length ORF including signal sequence (ss) and linker sequence (“long”), the ORF without the putative ss (based on the mature walnut sequence, “intermediate”), and the putative mature hazelnut sequence starting from the N ‐terminal sequence found in the purified natural protein (“short”).…”

Section: Discussionmentioning

confidence: 99%

Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut

Garino

Zuidmeer

Marsh

et al. 2010

Molecular Nutrition Food Res

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“…The spectrum displayed two negative bands, one at 208 and another at 222 nm, a positive band at 192 nm ( Figure 3a), and its deconvolution estimated 58% of α-helix, 12% of β-turns, 8% of β-sheets, and 22% of disordered structures. The cluster analysis indicated that AaCI-2S belonged to the α+β class of proteins, which presented a more pronounced band at 208 nm than the one at 222 nm [17], the typical secondary structure of members of the prolamin superfamily [18], similar to napin [19] and 2S albumin isolated from melon Momordica charantia [20].…”

Section: Secondary Structure Estimation and Intrinsic Fluorescence Emmentioning

confidence: 95%

Biotechnological Potential of Araucaria angustifolia Pine Nuts Extract and the Cysteine Protease Inhibitor AaCI-2S

Sallai

Salu

Silva-Lucca

et al. 2020

Plants

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Protease inhibitors are involved in the regulation of endogenous cysteine proteases during seed development and play a defensive role because of their ability to inhibit exogenous proteases such as those present in the digestive tracts of insects. Araucaria angustifolia seeds, which can be used in human and animal feed, were investigated for their potential for the development of agricultural biotechnology and in the field of human health. In the pine nuts extract, which blocked the activities of cysteine proteases, it was detected potent insecticidal activity against termites (Nasutitermes corniger) belonging to the most abundant termite genus in tropical regions. The cysteine inhibitor (AaCI-2S) was purified by ion-exchange, size exclusion, and reversed-phase chromatography. Its functional and structural stability was confirmed by spectroscopic and circular dichroism studies, and by detection of inhibitory activity at different temperatures and pH values. Besides having activity on cysteine proteases from C. maculatus digestive tract, AaCI-2S inhibited papain, bromelain, ficin, and cathepsin L and impaired cell proliferation in gastric and prostate cancer cell lines. These properties qualify A. angustifolia seeds as a protein source with value properties of natural insecticide and to contain a protease inhibitor with the potential to be a bioactive molecule on different cancer cells.

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In vitro refolded napin-like protein of Momordica charantia expressed in Escherichia coli displays properties of native napin

Cited by 14 publications

References 39 publications

RNase MC2: a new Momordica charantia ribonuclease that induces apoptosis in breast cancer cells associated with activation of MAPKs and induction of caspase pathways

RNase MC2: a new Momordica charantia ribonuclease that induces apoptosis in breast cancer cells associated with activation of MAPKs and induction of caspase pathways

Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut

Biotechnological Potential of Araucaria angustifolia Pine Nuts Extract and the Cysteine Protease Inhibitor AaCI-2S

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