Precursor of the Inactive 2S Seed Storage Protein from the Indian Mustard Brassica juncea Is a Novel Trypsin Inhibitor CHARACTERIZATION, POST-TRANSLATIONAL PROCESSING STUDIES, AND TRANSGENIC EXPRESSION TO DEVELOP INSECT-RESISTANT PLANTS
04 Oct 2002-Journal of Biological Chemistry (American Society for Biochemistry and Molecular Biology)-Vol. 277, Iss: 40, pp 37161-37168
TL;DR: A novel trypsin inhibitor from Indian mustard Brassica juncea that is unique in being the precursor of a 2S seed storage protein that can be used in transforming seed crops for protection to their vegetative parts and early seed stages, when insect damage is maximal.
About: This article is published in Journal of Biological Chemistry.The article was published on 2002-10-04 and is currently open access. It has received 53 citations till now. The article focuses on the topics: Trypsin inhibitor & Storage protein.
Citations
More filters
TL;DR: Initial indications from deployment of transgenics with insect resistance in diverse cropping systems in USA, Canada, Argentina, China, India, Australia, and South Africa suggest that single transgene products in standard cultivar backgrounds are not a recipe for sustainable pest management, and a much more complex approach may be needed.
Abstract: Transgenic resistance to insects has been demonstrated in plants expressing insecticidal genes such as δ -endotoxins from Bacillus thuringiensis (Bt), protease inhibitors, enzymes, secondary plant metabolites, and plant lectins. While transgenic plants with introduced Bt genes have been deployed in several crops on a global scale, the alternative genes have received considerably less attention. The protease inhibitor and lectin genes largely affect insect growth and development and, in most instances, do not result in insect mortality. The effective concentrations of these proteins are much greater than the Bt toxin proteins. Therefore, the potential of some of the alternative genes can only be realized by deploying them in combination with conventional host plant resistance and Bt genes. Genes conferring resistance to insects can also be deployed as multilines or synthetic varieties. Initial indications from deployment of transgenics with insect resistance in diverse cropping systems in USA, Canada, Arge...
146 citations
Cites background from "Precursor of the Inactive 2S Seed S..."
...A trypsin inhibitor from Indian mustard, Brassica juncea (BjTi), a precursor of a 2S seed storage protein, showed a soybean trypsin inhibitor active site like motif (GPFRI) at the expected processing site (Mandal et al., 2002)....
[...]
TL;DR: The spread, classification, and properties of plant proteins capable of inhibiting proteinases have been reviewed and data from the literature on the likely physiological functions of these inhibitors in plants are analyzed.
Abstract: The spread, classification, and properties of plant proteins capable of inhibiting proteinases have been reviewed. Data from the literature on the likely physiological functions of these inhibitors in plants are analyzed.
123 citations
TL;DR: A thorough assessment of the current literature suggests that, whereas the non-specific inhibitory effects of recombinant protease inhibitors in plant food webs could often be negligible and their 'unintended' pleiotropic effects in planta of potential agronomic value, the innocuity of these proteins might always remain an issue to be assessed empirically, on a case-by-case basis.
Abstract: Protease inhibitors are a promising complement to Bt toxins for the development of insect-resistant transgenic crops, but their limited specificity against proteolytic enzymes and the ubiquity of protease-dependent processes in living organisms raise questions about their eventual non-target effects in agroecosystems. After a brief overview of the main factors driving the impacts of insect-resistant transgenic crops on non-target organisms, the possible effects of protease inhibitors are discussed from a multitrophic perspective, taking into account not only the target herbivore proteases but also the proteases of other organisms found along the trophic chain, including the plant itself. Major progress has been achieved in recent years towards the design of highly potent broad-spectrum inhibitors and the field deployment of protease inhibitor-expressing transgenic plants resistant to major herbivore pests. A thorough assessment of the current literature suggests that, whereas the non-specific inhibitory effects of recombinant protease inhibitors in plant food webs could often be negligible and their 'unintended' pleiotropic effects in planta of potential agronomic value, the innocuity of these proteins might always remain an issue to be assessed empirically, on a case-by-case basis.
104 citations
Cites background from "Precursor of the Inactive 2S Seed S..."
...…Bovine spleen trypsin inhibitor Serine Helicoverpa armigera Christeller et al., 2002 Brassica juncea trypsin inhibitor Serine Spodoptera litura Mandal et al., 2002 Mustard trypsin inhibitor 2 Serine Spodoptera littoralis De Leo and Gallerani, 2002 Tobacco trypsin protease inhibitor Serine S.…...
[...]
TL;DR: This review will focus on the relevance of the structural-function relations of AMPs derived from plants and their proper use in applications for human health and agribusiness.
104 citations
Cites background from "Precursor of the Inactive 2S Seed S..."
...They can act in stabilizing oil-in-water mulsions [16,115], inhibiting proteolytic enzymes [92] and also erforming antibacterial or antifungal activities....
[...]
Journal Article•
TL;DR: The results suggested that VPE was responsible for cleaving Asn-Gln bonds of a single precursor, PV100, to produce multiple seed proteins, and it is likely that the Asn -Gln stretches not only provide cleavage sites for VPE but also produce aminopeptidase-resistant proteins.
Abstract: Precursor-accumulating vesicles mediate transport of the precursors of seed proteins to protein storage vacuoles in maturing pumpkin seeds. We isolated the precursor-accumulating vesicles and characterized a 100-kDa component (PV100) of the vesicles. Isolated cDNA for PV100 encoded a 97,310-Da protein that was composed of a hydrophobic signal peptide and the following three domains: an 11-kDa Cys-rich domain with four CXXXC motifs, a 34-kDa Arg/Glu-rich domain composed of six homologous repeats, and a 50-kDa vicilin-like domain. Both immunocytochemistry and immunoblots with anti-PV100 antibodies showed that <10-kDa proteins and the 50-kDa vicilin-like protein were accumulated in the vacuoles. To identify the mature proteins derived from PV100, soluble proteins of the vacuoles were separated, and their molecular structures were determined. Mass spectrometry and peptide sequencing showed that two Cys-rich peptides, three Arg/Glu-rich peptides, and the vicilin-like protein were produced by cleaving Asn-Gln bonds of PV100 and that all of these proteins had a pyroglutamate at their NH2 termini. To clarify the cleavage mechanism, in vitro processing of PV100 was performed with purified vacuolar processing enzyme (VPE). Taken together, these results suggested that VPE was responsible for cleaving Asn-Gln bonds of a single precursor, PV100, to produce multiple seed proteins. It is likely that the Asn-Gln stretches not only provide cleavage sites for VPE but also produce aminopeptidase-resistant proteins. We also found that the Cys-rich peptide functions as a trypsin inhibitor. Our findings suggested that PV100 is converted into different functional proteins, such as a proteinase inhibitor and a storage protein, in the vacuoles of seed cells.
93 citations
References
More filters
TL;DR: Inhibitors 0/ Carboxyl, Metallo and Sulfhydryl Proteinases 599 Inhibitors0/ Serine Proteinases 601 The Standard Mechanism 601 The nature of the reactive site 604 Multiple reactive sites on a single polypeptide chain 608 The Inhibitor Families
Abstract: PERSPECTIVES AND SUMMARY 593 INTRODUCTION 595 Limited Knowledge 0/ Physiological Function 595 Nomenclature Problems 596 0.2 MACROGLOBULINS 598 INHIBITORS WITH CLASS-SPECIFIC REACTIVE SITES 599 Inhibitors 0/ Carboxyl, Metallo and Sulfhydryl Proteinases 599 Inhibitors 0/ Serine Proteinases 601 The Standard Mechanism 601 The nature of the reactive site 604 Multiple reactive sites on a single polypeptide chain 608 The Inhibitor Families 610 The pancreatic trypsin inhibitor (Kunitzj family 610 Pancreatic secretory trypsin inhibitor (Kazal) family 613 Streptomyces subtilisin inhibitor family 616 Bowman-Berk inhibitor family : 616 Soybean trypsin inhibitor (Kunitzj family 617 Other inhibitor families 618 Inhibitors in mammalian blood plasma 619
2,139 citations
TL;DR: The inducible chemicals, which can also occur constitutively, include such complex sub stances as antibiotics, alkaloids, and terpenes, as well as proteins such as enzymes, enzyme inhibitors, and lectins, which have provided interesting systems for studying inducibles plant defense.
Abstract: For several decades researchers have been studying the biosynthesis and regulation in plants of specific chemicals associated with defense against pests and pathogens. For years many of these chemicals were first considered to be "secondary plant compounds" with unknown functions. Many, if not most, of these "secondary compounds" are now considered to be defensive chemicals that are acting either alone or in concert to contribute to the resistances of plants against insects or pathogens (86,87, 118, 144, 150). These defensive chemicals are found as either constitutive components in various plant tissues (118, 144) or are synthesized in response to attacking pests or pathogens (7, 32,39,45,50,60,63,64,66, 74, 120, 144, 151, 178, 182). The inducible chemicals, which can also occur constitutively, include such complex sub stances as antibiotics, alkaloids, and terpenes, as well as proteins such as enzymes, enzyme inhibitors, and lectins. These chemicals have provided interesting systems for studying inducible plant defense. In various plants, they can be newly synthesized in response to pest and/or pathogen attacks, which generate interand intracellular signals to activate genes that code for their production. Many of these systems, therefore, are being studied to help further our fundamental knowledge about the biochemistry of signal genera-
1,547 citations
TL;DR: All storage protein fractions are mixtures of components that exhibit polymorphism both within single genotypes and among genotypes of the same species, which arises from the presence of multigene families and, in some cases, proteolytic processing and glycosylation.
Abstract: in them. The presence of these groups may allow the plant to maintain high levels of storage protein synthesis despite variations in sulfur availability. The strict tissue specificity of seed storage protein synthesis contrasts with that of tuber storage proteins, which may be synthesized in vegetative tissues under unusual conditions (for example, in vitro or after removal of tubers) (Shewry, 1995). A second common property of seed storage proteins is their presence in the mature seed in discrete deposits called protein bodies, whose origin has been the subject of some dispute and may in fact vary both between and within species. Finally, all storage protein fractions are mixtures of components that exhibit polymorphism both within single genotypes and among genotypes of the same species. This polymorphism arises from the presence of multigene families and, in some cases, proteolytic processing and glycosylation.
959 citations
TL;DR: It is reported here that a gene encoding a cowpea trypsin inhibitor, which has been shown to give some measure of field resistance to insect pests, confers, when transferred to tobacco, enhanced resistance to this species' own herbivorous insect pests.
Abstract: A major goal of plant genetic engineering is the introduction of agronomically desirable phenotypic traits into crop plants in situations where conventional breeding methods have been unsuccessful. One such target is enhanced resistance to insect pests which, in view of the estimated production losses world-wide and the heavy costs of protective treatments, is very important. We report here that a gene encoding a cowpea trypsin inhibitor, which has been shown to give some measure of field resistance to insect pests1, confers, when transferred to tobacco, enhanced resistance to this species' own herbivorous insect pests.
931 citations
TL;DR: An efficient transformation system was developed for tomato by studying several factors that affect the rate of Agrobacterium mediated transformation of explant tissue and the tolerance phenotype segregated in a manner consistent with Mendelian inheritance.
Abstract: An efficient transformation system was developed for tomato by studying several factors that affect the rate of Agrobacterium mediated transformation of explant tissue. Cotyledons of tomato were co–cultivated with Agrobacterium tumefaciens harboring a binary vector with two neomycin phosphotransferase (NPT) II genes and a mutant aroA gene. Over 100 transgenic plants were regenerated and rooted on medium containing kanamycin and eighty percent of these plants exhibited both NPT II enzyme activity and produced the mutant aroA protein. Progeny of aroA positive plants were tolerant to glyphosate at concentrations of 0.84 kg active ingredient/ha. The tolerance phenotype segregated in a manner consistent with Mendelian inheritance.
516 citations