Analysis of secreted protease inhibitors after water stress in potato tubers

Ledoigt, Gérard; Griffaut, B.; Débiton, Eric; Vian, C.; Mustel, Ana; Evray, G.; Maurizis, Jean‐Claude; Madelmont, Jean‐Claude

doi:10.1016/j.ijbiomac.2006.03.016

Cited by 24 publications

(18 citation statements)

References 15 publications

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“…Protein fractions, approximately eight-fold concentrated, were called S2. Protein content was assayed according to Bradford [25] or using NanoOrange protein assay kit according to themolecular weights on sephacrylamide (S300-HR, Sigma) column as previously described [3]. Polypeptides were eluted with phosphate buffer solution (0.2 M, pH 7.5) and fractions of 3 ml or 1 ml were then recovered.…”

Section: Isolation and Purification Of The Cytotoxic Agentmentioning

confidence: 99%

“…Two-dimension isoelectrofocusing electrophoresis (2-D IEF PAGE) was performed according to O'Farrell [31]. Non-equilibrium pH gradient electrophoresis was done with fraction of secreted patatin-like proteins previously purified by PAGE in non-denaturing conditions [3]. First dimension was done in polyacrylamide gel (6.4% acrylamide) containing ampholines (pH 3.5 -10 (250 µl) or 5 -7 (450 µl): Pharmacia Biotech), at 4˚C, for 16 h (900 V); electrophoresis was realized using 0.02 M NaOH solution in cathode tank and 0.01 M H 3 PO 4 solution at anode.…”

Section: D-or 2d-sds-polyacrylamide Gel Electrophoresis (Sds-pages)mentioning

confidence: 99%

“…Non-denaturing PAGE of secretion products revealed three silver nitrate stained bands, two minor ones around 180 kDa (band A) and 40 kDa (band C) and a major band at 95 kDa (band B) [3]. Using SDS-PAGE, we previously described two sets of polypeptides which comigrate in band B: a major polypeptide (MW 35 -40 kDa) and a smaller one (around 16 kDa) containing Kunitz-type protease inhibitors [3].…”

Section: Secreted Protein Characterisationmentioning

confidence: 99%

“…Variations of sequences mainly occurred at the level of first and third N-terminal amino acid. A protease inhibitor (spot B5) often is associated to patatin complex (Figure 1) [3]. 2D IEF spots were removed from gel and analysed by MALDI-TOF method ( Table 1).…”

Section: Pla 2 Activity Assaysmentioning

confidence: 99%

“…This protein alone or possibly associated with a co-factor was shown to induce cytotoxicity in plant and animal tumour cells, preserving non-tumour cells in both cases. In potato (Solanum tuberosum), after such a stress condition, tuber parenchyma was shown to secrete new types of Kunitz-like protease inhibitors closely associated with another unknown polypeptide of approximately 40 kDa [3].…”

Section: Introductionmentioning

confidence: 99%

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Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells

Griffaut¹,

Débiton²,

Galmier³

et al. 2013

ABC

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Plants can respond to traumatism by synthesis and secretion of defence molecules. Wound-healing and desiccation stress of pieces of potato tuber parenchyma (Solanum tuberosum) promoted the secretion of a patatin-like phospholipase A 2 , PLA 2 (EC.3.1.1.4.) that displayed cytotoxic activity against tumour cells. The potato secretion product, an oligomeric form of patatin-like protein, was shown to contain several isoforms of PLA 2 polypeptides and to be associated with other proteins, including Kunitz-type protease inhibitors. Patatin-like protein secretion was inhibited by vanadate. Secreted patatin-like proteins displayed specific features, such as extracellular function and low molecular weights, mainly 36 to 40 kDa. The 36-kDa polypeptide sequence was related to iPLA 2 α. Polypeptide spots of secreted patatin-like protein exhibited a nucleotide-binding consensus motif, GGGIKG that has been described in iPLA 2 gene family. The cytotoxic agent caused cell death of plant crown-gall induced by Agrobacterium tumefaciens, and inhibited B16 cell proliferation, but at the same concentration did not display any toxicity against non-transformed cells. PLA 2 enzyme activity was required for cytotoxicity against B16 melanoma cells. A model for such a specific activity against tumour cells is discussed in connection with asymmetric phospholipid patterns of cell membranes. In conclusion, secreted patatin-like PLA 2 (e-patatin) may represent a novel therapeutic target for the development of new agents against cancer.

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Section: Isolation and Purification Of The Cytotoxic Agentmentioning

confidence: 99%

Section: D-or 2d-sds-polyacrylamide Gel Electrophoresis (Sds-pages)mentioning

confidence: 99%

Section: Secreted Protein Characterisationmentioning

confidence: 99%

Section: Pla 2 Activity Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells

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Untitled

Nawaz

Sufyan²,

Tayyab³

et al. 2022

Advances in Integrated Pest Management Technology

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Identification and expression analysis of salt-responsive genes using a comparative microarray approach in Salix matsudana

et al. 2014

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Salt stress exerts negative effects on plant growth, development and yields, with roots being the primary site of both perception and damage. Salix matsudana (Chinese willow) is tolerant of high salinity. However, genes associated with this trait were rarely characterized. Therefore, we first performed salt-stress treatment on S. matsudana plants, then identified differentially expressed genes by comparison of salt-treated roots and untreated controls using microarray analysis. A total of 403 salt-responsive genes were identified, of which 239 were repressed and 164 were up-regulated. Functional classification analysis revealed that these genes belonged to families encoding proteins involved in metabolism, regulation of transcription, signal transduction, hormone responses, abiotic stress responses, and other processes related to growth and development. This suggested that when S. matsudana was confronted with salt stress, coordinated adjustments are made to physiological and biochemical processes, which would then allow more resources to be allocated to protective mechanisms to avoid salt injury. The expression patterns of representative genes were further validated and the diversity of the temporal profiles indicated that a combination of several genes and the initiation of diverse pathways performed functions in S. matsudana salt tolerance. This work represents the first study employing microarrays to investigate salt tolerance in S. matsudana. The data presented herein enhance our understanding of the molecular mechanisms of S. matsudana responses to salinity stress and lay the groundwork for genetic engineering strategies to improve stress tolerance of agronomically important species.

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Analysis of secreted protease inhibitors after water stress in potato tubers

Cited by 24 publications

References 15 publications

Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells

Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells

Untitled

Identification and expression analysis of salt-responsive genes using a comparative microarray approach in Salix matsudana

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Analysis of secreted protease inhibitors after water stress in potato tubers

Cited by 24 publications

References 15 publications

Patatin-like PLA&lt;sub&gt;2&lt;/sub&gt; with cytotoxicity against mammalian and plant tumour cells

Patatin-like PLA&lt;sub&gt;2&lt;/sub&gt; with cytotoxicity against mammalian and plant tumour cells

Untitled

Identification and expression analysis of salt-responsive genes using a comparative microarray approach in Salix matsudana

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Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells

Patatin-like PLA<sub>2</sub> with cytotoxicity against mammalian and plant tumour cells