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1.
Isolation and Partial Characterization of a Seed Lectin from Tepary Bean that Delays Bruchid Beetle Development 总被引:2,自引:1,他引:1
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Four isolectin forms of a seed lectin from mature seed of tepary bean (Phaseolus acutifolius) were isolated using solubility fractionation, affinity chromatography, and high performance liquid chromatography. The subunits are polypeptides with an apparent molecular mass of 30,000 daltons. The 30 kilodalton subunits are produced starting approximately 13 days after flowering and subsequently comprise a major fraction of the proteins found in the mature seed. The amino terminus of each isolectin fraction was determined to be highly homologous with that of the subunits of common bean (Phaseolus vulgaris L.) phytohemagglutinin (PHA). The tepary isolectin cross-reacts with both erythroagglutinating and leucoagglutinating subunits of PHA antibodies, although differential cross-reactivity was noted. A seed protein fraction enriched in tepary bean lectin was found to be toxic to bean bruchid beetles (Acanthoscelides obtectus), when incorporated into their diets at incremental concentrations from (1-5% w/w) above that of PHA concentrations in mature seeds of the susceptible common bean variety “Red Kidney.” 相似文献
2.
《Phytochemistry》1987,26(2):335-343
Phaseolin, the major storage protein of the French bean Phaseolus vulgaris cv. Tendergreen, has been isolated and purified by either ion-exchange chromatography or reversed-phase HPLC. These purification procedures were used to fractionate the native protein aggregate into its characteristic subunit components. Amino-terminal sequence analysis was performed on the intact peptide subunits. Native phaseolin was chemically cleaved at a unique tryptophan residue which is proximal to the N-terminal region of the protein with BNPS-skatole and the resulting peptide fragments were isolated via reversed-phase HPLC. Chemical and enzymatic sequence results obtained from these peptide fragments are in full agreement with the results obtained for the full length peptide subunits. These N-terminal analyses show that the signal peptide cleavage process is somewhat random resulting in the phaseolin polypeptides having possibly three or four different N-termini. Native phaseolin and purified subunits were chemically deglycosylated with trifluoromethanesulphonic acid in the presence of an anisole scavenger. One-dimensional SDS-PAGE analysis of the deglycosylated products show that differential glycosylation is largely responsible for much of the observed molecular weight heterogeneity found among phaseolin polypeptides. 相似文献
3.
Proteinase A-like enzyme from germinated kidney bean seeds. Its action on phaseolin and vicilin 总被引:3,自引:0,他引:3
Vitalyi Rotari Vitalyi Senyuk Christian Horstmann Antonina Jivotovskaya Iosif Vaintraub 《Physiologia plantarum》1997,100(1):171-177
A cysteine proteinase that possibly participates in the degradation of phaseolin, the main storage protein of kidney bean ( Phaseolus vulgaris L. cv. Moldavian) was isolated from germinating kidney bean seeds and partially characterized. According to its properties it may be classified as a member of a group of homologous cysteine proteinases A, also present in germinating seeds of a number of other plants. The proteinase of this group hydrolyze storage proteins to short peptides. Similarly, the kidney bean proteinase hydrolyzes vicilin, the reserve protein of vetch ( Vicia sativa ). However, its action on phaseolin is limited to the cleavage of subunits into two approximately equal parts and to the splitting off a small number of short peptides. An explanation of phaseolin resistance to the action of this proteinase is proposed on the basis of the differences of its structure from that of other homologous 7S proteins. 相似文献
4.
The structure of canavalin, a jack bean (Canavalis ensiformis) protein homologous to phaseolin, the major seed storage protein of Phaseolus vulgaris, has been investigated by x-ray crystallography and found to be a hexamer composed of three identical pairs of similar but nonidentical subunits related by a perfect 3-fold axis and pseudo dyad axes (strict C3 and pseudo D3). One member of each pair of subunits is derived from the amino terminal half of a precursor polypeptide of molecular weight 49,000 and the other from its carboxy terminal half. Thus, the crystallographic evidence indicates that the precursor polypeptide is a tandem duplicate and is structurally redundant (McPherson A. 1982 J Biol Chem 255: 10472). A number of physical and chemical properties of the protein in both the uncleaved and the cleaved form were investigated. These included the native molecular weights, amino acid analyses, number of exposed sulfhydryl groups, carbohydrate content, metal ion analysis, crystallization behavior, and the fate of the protein in developing seeds. It was also found that the purified precursor protein possesses a substantial level of α-d-mannosidase activity and seems to share a number of other physical and chemical properties with that enzyme. 相似文献
5.
Ademir Sérgio Ferreira de Araujo Angela Celis de Almeida Lopes Jorge C. Berny Mier y Teran Antonia Palkovic Paul Gepts 《Symbiosis (Philadelphia, Pa.)》2017,73(1):7-14
Phaseolus lunatus is the second economically most important species of the genus Phaseolus. It carries out N fixation through symbiosis with rhizobia. However, it is unclear whether P. lunatus can nodulate with native rhizobia from soils where this legume is not native or was not cultivated previously. Thus, this study assessed the ability of 14 geographically distant lima bean genotypes to nodulate with rhizobia from three California agricultural soils: without a history of legumes or P. lunatus cultivation, with a history of legumes as a cover crop, and with a history of P. lunatus cultivation. Nodulation only occurred on genotypes grown in the soil with a history of P. lunatus planting. The analysis of variance of nodulation traits showed that the genotype effect was highly significant in all the traits measured. Shoot biomass had a higher correlation with nodule size and nodule weight than with nodule number. In addition, shoot biomass and leaf N content were positively correlated with nodule coloration and with nodule position close to the main root of the plant. This study suggests that agricultural soils from California do not appear to have native rhizobia able to nodulate P. lunatus, which suggests the need to inoculate, at least initially, the seeds at planting in order to establish the population of rhizobia. Also, geographically distant lima bean genotypes have different responses to nodulating bacteria and it suggests that future studies to test these genotypes across different environments should be pursued. 相似文献
6.
Peptide Mapping Reveals Considerable Sequence Homology among the Three Polypeptide Subunits of G1 Storage Protein from French Bean Seed 总被引:4,自引:0,他引:4
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The major storage protein, G1 globulin, of bean (cv. Tendergreen) seeds was subjected to limited proteolysis with trypsin, chymotrypsin, papain, proteinase K, and protease V8 and to cleavage with cyanogen bromide and 2-(2-nitrophenylsulfanyl)-3-methyl-3′bromoindolenine. Mapping of peptides separated from each of the three G1 subunits by polyacrylamide gel electrophoresis revealed that many proteolytic cleavage sites were present at similar positions on the subunits. Evidence was adduced that the G1 subunits are homologous in amino acid sequence for about 61% of their length. The remaining region (possibly COOH-terminal) of the subunits appears to be heterologous, with the α subunit bearing an additional methionine residue. 相似文献
7.
Microheterogeneity of globulin-1 storage protein from French bean with isoelectrofocusing 总被引:3,自引:1,他引:2
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The major storage protein fraction, globulin-1 protein, of French bean (Phaseolus vulgaris L.) was analyzed by two-dimensional electrophoresis. The protein pattern suggested a more complex system for globulin-1 protein than the model of three polypeptides, α, β, and γ, differing in molecular weight. Isoelectrofocusing analyses of the individual proteins showed that each exhibited charge microheterogeneity over a similar pH range. Isoelectrofocusing banding patterns may help to understand the relationships between the globulin-1 polypeptide subunits. 相似文献
8.
Paul Gepts 《Economic botany》1990,44(3):28-38
The genusPhaseolus (Fabaceae) consists of some 50 species, all of which are distributed in the Americas. Four of these contain cultigens.P. vulgaris (common bean),P. lunatus (lima bean),P. acutifolius (tepary bean),P. coccineus subsp.coccineus (runner bean); andP. coccineus subsp.polyanthus (no English vernacular name). Biochemical markers—phaseolin seed storage protein and isozymes—have provided new evidence on the organization of the first three species. Domestication has possibly caused a strong reduction in genetic diversity inP. vulgaris andP. acutifolius. BothP. vulgaris andP. lunatus cultivars result from at least two independent domestications, in Mesoamerica and in the Andes. These two species consist of two gene pools, each of which includes wild ancestors and their respective cultivated descendants. Our findings suggest the need for additional emphasis on genetic conservation of wild ancestors and their use in breeding programs and for a comparison of inter-gene pool vs. intra-gene pool crosses in breeding programs. 相似文献
9.
Purification and characterization of two alpha-amylase inhibitors from seeds of tepary bean (Phaseolus acutifolius A. Gray) 总被引:2,自引:0,他引:2
Two proteinaceous alpha-amylase inhibitors termed alphaAI-Pa1 and alphaAI-Pa2 were purified from seeds of a cultivated tepary bean (Phaseolus acutifolius A. Gray, cv. PI311897). The two inhibitors differed in their specificity towards alpha-amylases of insect pests such as bruchids, although neither showed any inhibitory activity against alpha-amylases of mammalian, bacterial or fungal origin. AlphaAI-Pa2 resembles two common bean inhibitors, alphaAI-1 and alphaAI-2, in several characteristics such as N-terminal amino acid sequences and oligomeric structure being composed of alpha and beta subunits. In contrast alphaAI-Pa1 is composed of a single glycopolypeptide with a molecular mass of 35 kDa, and its N-terminal amino acid sequence resembled that of seed lectins in tepary bean and common bean. The information on the two tepary bean alpha-amylase inhibitors may be useful not only for providing insight into critical structure for the specificity towards different alpha-amylase enzymes but also for enhancing insect resistance in crops. 相似文献
10.
Inès Karmous Jaouani Khadija Abdelilah Chaoui Ezzedine El Ferjani 《Physiology and Molecular Biology of Plants》2012,18(4):337-343
The changes in the protease activities of bean cotyledons were investigated in response to copper stress. Assays using synthetic substrates and specific protease inhibitors followed by activity measurements and electrophoresis analysis allowed to study the classes of enzymes involved in the storage protein mobilization during the germination of bean (Phaseolus vulgaris L) seeds, and then identify which ones were affected in the presence of 200 μM CuCl2 in the imbibition medium. Copper treatment affected embryo growth and total protease activity. The results of SDS-gelatin-PAGE show that Cu excess led to a decrease in protease activity of 45 to 66 kDa. Moreover, cysteine-, aspartic- and metallo-protease activities were markedly lowered under copper stress, while serine-protease one was enhanced as well as its activity dependent abundance in comparison with control. However, the relative distribution of major cysteine protease in H2O-germinated seeds was significantly diminished after Cu exposure. Thus, copper excess can disturb the nitrogen freeing from reserve tissues at enzymatic level; differential responses of protease classes are discussed, notably, cysteine protease in the way of storage protein mobilization and serine protease in protective mechanism one. 相似文献
11.
O. P. Sehgal 《Plant cell reports》1992,12(1):55-59
Tobacco ringspot virus (TRSV) induces circular, darkbrown local lesions on primary leaves of lima bean (Phaseolus lunatus cv Nemagreen) with a concomitant production of three basic and three acidic pathogenesisrelated (PR) proteins. The three basic proteins are: a 21 kDa protein related serologically to Pinto bean PR-4d and tobacco PR-5 proteins; a 36 kDa glucanase that is related to tobacco PR-2; and, a 31 kDa chitinase related serologically to ethylene-induced bean chitinase. The three acidic 18 kDa lima bean PR proteins are serologically similar and probably are charged isomers of the same protein. The 21 kDa basic protein and the 18 kDa acidic protein accumulated preferentially at the lesion center while the 31 kDa chitinase and TRSV were distributed evenly throughout the necrotic area. In green tissue immediately surrounding a lesion, the amounts of PR proteins were comparable to or lower than those in the necrotic area, and virions were not detected. This mode of spatial distribution indicates that lima bean PR proteins are not involved in TRSV localization, and is consistent with other observations that PR proteins play no direct role in restricting viral spread. 相似文献
12.
The acidic and basic subunits of glycinin—the major storage protein of soybean seeds (Glycine max)—were isolated by a simple ion exchange chromatography method involving a two-step pH change of the elution buffer. Abnormal behaviour of the subunits in dodecyl sulfate polyacrylamide gel electrophoresis is suggested. 相似文献
13.
Angelo Santino Antonia Gallo Antonella Leone Palmiro Poltronieri Giuseppe Zacheo 《Physiologia plantarum》1998,104(3):293-298
A proteinaceous inhibitor of papain was purified to apparent homogeneity from mature seeds of common bean ( Phaseolus vulgaris L.). After four chromatographic steps, the papain inhibitor was purified 219‐fold with 12% recovery. On the basis of papain inhibitory activity, cystatins have been estimated to account for about 0.1% of the total protein content of mature common bean seeds. The purified protein, as other plant cystatins, is an acidic protein, heat stable and insensitive to reducing agents. Its molecular mass is about 37 kDa as judged by size exclusion chromatography and SDS‐PAGE. Moreover it is immunologically related to oryzacystatins, since it is recognised by a specific oryzacystatin I antiserum. Based on its biochemical properties the papain inhibitor described here belongs to the phytocystatin family. Papain inhibitory assays carried out during seed development showed that bean cystatin is active since early maturation stages. Our results suggest that, in common bean seed, cysteine proteinase inhibitors are important during seed development with a putative role in the control and regulation of endogenous thiol protease activity. 相似文献
14.
S. N. Kulikov S. N. Chirkov A. V. Il’ina S. A. Lopatin V. P. Varlamov 《Applied Biochemistry and Microbiology》2006,42(2):200-203
The effect of the molecular weight of chitosan on its ability to suppress systemic infection of bean mild mosaic virus in bean (Phaseolus vulgaris L.) plants was studied. The enzymatic hydrolysate of low-molecular-weight chitosan was successively fractionated by ultrafiltration through membranes with decreasing pore size. In total, four chitosan fractions with a weight-average molecular weight varying from 1.2 to 40.4 kDa were obtained. It was shown that the treatments of bean plants with these fractions (chitosan concentration, 10 or 100 μg/ml) inhibited virus accumulation and systemic propagation. The degree of chitosan-induced antiviral resistance increased as the molecular weight of chitosan decreased. The monomers comprising the chitosan molecule—glucosamine and N-acetylglucosamine—exhibited no antiviral activity. 相似文献
15.
Zakharov A Carchilan M Stepurina T Rotari V Wilson K Vaintraub I 《Journal of experimental botany》2004,55(406):2241-2249
Two types of cysteine proteases, low-specificity enzymes from the papain family and Asn-specific from the legumain family are generally considered to be the major endopeptidases responsible for the degradation of seed storage proteins during early seedling growth. The action of the corresponding enzymes (CPPh1 and LLP, respectively) from common bean (Phaseolus vulgaris L.) on phaseolin (the common bean storage protein), and on the homologous soybean (Glycine max (L.) Merrill) storage protein, beta-conglycinin, was studied. Under the action of LLP, proteolysis of phaseolin was limited to cleavage of its interdomain linker. No cleavage of the interdomain linker occurred in beta-conglycinin with LLP. LLP action was restricted to splitting off the disordered N-terminal extensions of alpha and alpha' subunits. No extensive hydrolysis (degradation to short TCA-soluble peptides) of either protein occurred under the action of LLP. CPPh1 cleaved the phaseolin subunits into roughly half-sized fragments at the onset of proteolysis. The cleavage was accompanied by a small (8-10%) decrease of protein. No decrease of protein occurred with further incubation. Thus the two most active proteinases detected in common bean seedlings individually were incapable of the extensive degradation of phaseolin. Extensive hydrolysis of phaseolin was only achieved by the consecutive action of LLP and CPPh1. Similar cleavages occurred during the action of CPPh1 on beta-conglycinin. However, by contrast with phaseolin, CPPh1 by itself accomplished the extensive hydrolysis of beta-conglycinin. The differences in the course of proteolysis of the proteins studied were determined by their structural peculiarities. 相似文献
16.
Josef Kloz 《Biologia Plantarum》1962,4(2):85-90
Byla provedena kvantitativni a kvalitativní analysa bílkovinných znak? z děloh v semenech a z hypokotyl? — ko?ínk? (primárního ko?ene) naklí? enÝch semen těchto druh?:Phaseolus vulgaris L.,Phaseolus coccineus L.,Phaseolus lunatus L.,Phaseolus aureus ROXB. s těmito vÝsledky:
- 1.Fazeolin témě? shodnÝ je v dělohách druh?Phaseolus vulgaris L aPhaseolus coccines L.; u druh?Phaseolus lunatus L. aPhaseolus aureus Roxb. chybí. 相似文献
17.
A cysteine endopeptidase isolated from castor bean endosperm microbodies processes the glyoxysomal malate dehydrogenase precursor protein. 总被引:2,自引:1,他引:1
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A plant cysteine endopeptidase with a molecular mass of 35 kD was purified from microbodies of germinating castor bean (Ricinus communis) endosperm by virtue of its capacity to specifically process the glyoxysomal malate dehydrogenase precursor protein to the mature subunit in vitro. Processing of the glyoxysomal malate dehydrogenase precursor occurs sequentially in three steps, the first intermediate resulting from cleavage after arginine-13 within the presequence and the second from cleavage after arginine-33. The endopeptidase is unable to remove the presequences of prethiolases from rape (Brassica napus) glyoxysomes and rat peroxisomes at the expected cleavage site. Protein sequence analysis of N-terminal and internal peptides revealed high identity to the mature papain-type cysteine endopeptidases from cotyledons of germinating mung bean (Vigna mungo) and French bean (Phaseolus vulgaris) seeds. These endopeptidases are synthesized with an extended pre-/prosequence at the N terminus and have been considered to be processed in the endoplasmic reticulum and targeted to protein-storing vacuoles. 相似文献
18.
The protease that degrades the beta subunit of the soybean (Glycine max (L.) Merrill) storage protein beta-conglycinin was purified from the cotyledons of seedlings grown for 12 days. The enzyme was named protease C2 because it is the second enzyme to cleave the beta-conglycinin storage protein, the first (protease C1) being one that degrades only the alpha' and alpha subunits of the storage protein to products similar in size and sequence to the remaining intact beta subunit. Protease C2 activity is not evident in vivo until 4 days after imbibition of the seed. The 31 kDa enzyme is a cysteine protease with a pH optimum with beta-conglycinin as substrate of 5.5. The action of protease C2 on native beta-conglycinin produces a set of large fragments (52-46 kDa in size) and small fragments (29-25 kDa). This is consistent with cleavage of all beta-conglycinin subunits at the region linking their N- and C-domains. Protease C2 also cleaves phaseolin, the Phaseolus vulgaris vicilin homologous to beta-conglycinin, to fragments in the 25-28 kDa range. N-Terminal sequences of isolated beta-conglycinin and phaseolin products show that protease C2 cleaves at a bond within a very mobile surface loop connecting the two compact structural domains of each subunit. The protease C2 cleavage specificity appears to be dictated by the substrate's three-dimensional structure rather than a specificity for a particular amino acid or sequence. 相似文献
19.
A non-specific lipid transfer protein with antifungal and antibacterial activities from the mung bean 总被引:11,自引:0,他引:11
A non-specific lipid transfer peptide (nsLTP) with antimicrobial activity was isolated from the mung bean (Phaseolus mungo) seeds. The procedure entailed aqueous extraction, ion exchange chromatography on CM-Sephadex and high performance liquid chromatography (HPLC) on POROS-HS-20. The peptide exhibited a molecular mass of 9.03 kDa in mass spectrometry. It exerted antifungal action toward Fusarium solani, Fusarium oxysporum, Pythium aphanidermatum and Sclerotium rolfsii, and antibacterial action against Staphylococcus aureus but not against Salmonella typhimurium. The lipid binding of this peptide was very similar to that of a previously described lipid transfer protein extracted from wheat seeds and maize seeds, indicating that it possessed lipid transfer activity. The present findings add to the scarcity of the literature on leguminous nsLTPs. 相似文献
20.
Lucia Lioi 《Biologia Plantarum》1987,29(1):49-53
Relationships among storage proteins in seeds from cultivars and primitive accessions of the four economically most important
species ofPhaseolus — P. vulgaris, P. coccineus, P. acutifolius andP. lunatus — were studied.
Analysis of SDS-polyacrylamide gel electrophoretic patterns of storage seed proteins revealed common characteristics in the
major groups of polypeptides forP. vulgaris, P. coccineus andP. acutifolius, while clear differences existed between thesePhaseolus species and P.lunatus. 相似文献