Jasmonic acid inducible aspartic proteinase inhibitors from potato

Kreft, Samo; Ravnikar, Maja; Meško, Pika; Pungerčar, Jože; Umek, Andrej; Kregar, I.; Štrukelj, Borut

doi:10.1016/s0031-9422(96)00668-1

Cited by 31 publications

(15 citation statements)

References 22 publications

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“…Sequence analysis revealed that the cDNA corresponds to a possible CDI (Werner et al, 1993). In fact, the JIP21-deduced amino acid sequence presents an elevated identity with the family of CDIs described in potato (Mares et al, 1989;Ritonja et al, 1990;Strukelj et al, 1990;Hildmann et al, 1992;Maganja et al, 1992;Hannapel, 1993;Herbers et al, 1994;Ishikawa et al, 1994a;Kreft et al, 1997). Apart from this large family in potato, proteinaceous aspartic PINs are uncommon and are described in only a few plant species, yeast (Schu and Wolf, 1991), and the nematode Ascaris lumbricoides (AbuErreish and Peanasky, 1974).…”

Section: Discussionmentioning

confidence: 99%

“…Since then, up to 15 isoforms have been found in potato and they have been classified as CDIs exclusively on the basis of sequence analysis. Potato CDIs constitutively accumulate in tubers and flower buds and are also induced by wounding in potato leaves Hildmann et al, 1992;Maganja et al, 1992;Hannapel, 1993;Herbers et al, 1994;Ishikawa et al, 1994a;Kreft et al, 1997).…”

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confidence: 99%

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A Novel Function for the Cathepsin D Inhibitor in Tomato

2006

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Proteinaceous aspartic proteinase inhibitors are rare in nature and are described in only a few plant species. One of them corresponds to a family of cathepsin D inhibitors (CDIs) described in potato (Solanum tuberosum), involving up to 15 isoforms with a high sequence similarity. In this work, we describe a tomato (Solanum lycopersicum) wound-inducible protein called jasmonic-induced protein 21 (JIP21). Sequence analysis of its cDNA predicted a putative function as a CDI. The JIP21 gene, whose protein has been demonstrated to be glycosylated, is constitutively expressed in flowers, stem, and fruit, and is inducible to high levels by wounding and methyl jasmonate in leaves of tomato plants. The genomic sequence of JIP21 shows that the gene is intronless and reveals the presence of both a methyl jasmonate box (TGACT) and a G-box (CACGT) in the promoter. In contrast to the presumed role of JIP21 based on sequence analysis, a detailed biochemical characterization of the purified protein uncovers a different function as a strong chymotrypsin inhibitor, which questions the previously predicted inhibitory activity against aspartic proteinases. Moreover, Egyptian cotton worm (Spodoptera littoralis) larvae fed on transgenic tomato plants overexpressing JIP21 present an increase in mortality and a delay in growth when compared with larvae fed on wild-type plants. These larvae belong to the Lepidoptera family whose main digestive enzymes have been described as being Ser proteases. All these results support the notion that tomato JIP21 should be considered as a chymotrypsin inhibitor belonging to the Ser proteinase inhibitors rather than a CDI. Therefore, we propose to name this protein tomato chymotrypsin inhibitor 21 (TCI21).

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

A Novel Function for the Cathepsin D Inhibitor in Tomato

2006

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“…Oryzacystatin I (OCI) (Abe et al, 1987) was expressed as s glutathione S-transferase (GST) fusion protein in Escherichia coli strain BL21 (Stratagene) as previously described (Michaud et al, 1994). Aspartyl proteinase inhibitor (API) was purified from potato as previously described (Kreft et al, 1997) and consisted of several isoforms (Ritonja et al, 1990;Barlic-Maganja et al, 1992;Strukelj et al 1992Strukelj et al , 1995Kreft et al, 1997) of potato cathepsin D inhibitor (PDI) (Keilova and Tomasek, 1976b).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors

Wilhite

Elden

Brzin

et al. 2000

Insect Biochemistry and Molecular Biology

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"Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors" (2000). AbstractProteolytic activities in alfalfa weevil (Hypera postica) larval midguts have been characterized. Effects of pH, thiol activators, low-molecular weight inhibitors, and proteinase inhibitors (PIs) on general substrate hydrolysis by midgut extracts were determined. Hemoglobinolytic activity was highest in the acidic to mildly acidic pH range, but was maximal at pH 3.5. Addition of thiolactivators dithiothreitol (DTT), 2-mercaptoethanol (2-ME), or l-cysteine had little effect on hemoglobin hydrolysis at pH 3.5, but enhanced azocaseinolytic activity two to three-fold at pH 5.0. The broad cysteine PI E-64 reduced azocaseinolytic activity by 64% or 42% at pH 5 in the presence or absence of 5 mM l-cysteine, respectively. Inhibition by diazomethyl ketones, Z-Phe-Phe-CHN 2 and Z-Phe-Ala-CHN 2 , suggest that cathepsins L and B are present and comprise approximately 70% and 30% of the cysteine proteolytic activity, respectively. An aspartyl proteinase component was identified using pepstatin A, which inhibited 32% (pH 3.5, hemoglobin) and 50% (pH 5, azocasein) of total proteolytic activity. This activity was completely inhibited by an aspartyl proteinase inhibitor from potato (API), and is consistent with the action of a cathepsin D-like enzyme. Hence, genes encoding PIs with specificity toward cathepsins L, B and D could potentially be effective for control of alfalfa weevil using transgenic plants.

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“…The recombinant plant cyprosin was not affected (IC 50 Ͼ Ͼ 2,000 nM) by a plant derived protein isolated from potatoes, which is a potent inhibitor (K i ϳ4 nM) of cathepsin D (32). Similarly, the naturally occurring protein inhibitors of pepsin/ cathepsin E and yeast proteinase A from the parasitic worm A. lumbricoides (33) and S. cerevisiae (34), respectively, did not inhibit the recombinant cyprosin to any significant extent (IC 50 Ͼ Ͼ 2,000 nM).…”

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confidence: 98%

Processing, Activity, and Inhibition of Recombinant Cyprosin, an Aspartic Proteinase from Cardoon (Cynara cardunculus)

White

Cordeiro²,

Arnold

et al. 1999

Journal of Biological Chemistry

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The cDNA encoding the precursor of an aspartic proteinase from the flowers of the cardoon, Cynara cardunculus, was expressed in Pichia pastoris, and the recombinant, mature cyprosin that accumulated in the culture medium was purified and characterized. The resultant mixture of microheterogeneous forms was shown to consist of glycosylated heavy chains (34 or 32 kDa) plus associated light chains with molecular weights in the region of 14,000 -18,000, resulting from excision of most, but not all, of the 104 residues contributed by the unique region known as the plant specific insert. SDS-polyacrylamide gel electrophoresis under non-reducing conditions indicated that disulfide bonding held the heavy and light chains together in the heterodimeric enzyme forms. In contrast, when a construct was expressed in which the nucleotides encoding the 104 residues of the plant specific insert were deleted, the inactive, unprocessed precursor form (procyprosin) accumulated, indicating that the plant-specific insert has a role in ensuring that the nascent polypeptide is folded properly and rendered capable of being activated to generate mature, active proteinase. Kinetic parameters were derived for the hydrolysis of a synthetic peptide substrate by wildtype, recombinant cyprosin at a variety of pH and temperature values and the subsite requirements of the enzyme were mapped using a systematic series of synthetic inhibitors. The significance is discussed of the susceptibility of cyprosin to inhibitors of human immunodeficiency virus proteinase and particularly of renin, some of which were found to have subnanomolar potencies against the plant enzyme.

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Jasmonic acid inducible aspartic proteinase inhibitors from potato

Cited by 31 publications

References 22 publications

A Novel Function for the Cathepsin D Inhibitor in Tomato

A Novel Function for the Cathepsin D Inhibitor in Tomato

Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors

Processing, Activity, and Inhibition of Recombinant Cyprosin, an Aspartic Proteinase from Cardoon (Cynara cardunculus)

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