Effects of sodium dodecyl sulfate and other dissociating reagents on the globulins of peas

Grant, D.R.; Lawrence, John M.

doi:10.1016/0003-9861(64)90441-2

Cited by 46 publications

(17 citation statements)

References 27 publications

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“…The elution profile from the DEAE-cellulose column suggested to us that peak III and IV may correspond to vicilin and legumin, the two main storage proteins found in the various species of Leguminosae, because similar elution profiles have been obtained for the storage proteins of peas (8) and vetch (26). To confirm this idea, we determined the sedimentation constants of these two major protein fractions.…”

mentioning

confidence: 76%

“…9). Calculations based on an S value of 4.15 for sheep hemoglobin indicated S values of 8.0 for peak III (identical to B) and 11.3 for peak IV (identical to A). Peaks III and IV, therefore, probably correspond respectively to vicilin and legumin as identified by Danielsson (6).…”

mentioning

confidence: 92%

“…To determine whether or not any of the subunits may have been due to contamination from Peak IV, the protein in this peak was also concentrated and subjected to sodium dodecyl sulfate-acrylamide gel electrophore- Table I for molecular weights of these subunits. (3,8,10, and 32). However, no two reports agree, indicating that vicilin may be a heterogeneous protein and that there may be considerable variation from species to species.…”

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confidence: 99%

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Isolation and Characterization of Glucosamine-containing Storage Glycoproteins from the Cotyledons of Phaseolus aureus

Ericson¹,

Chrispeels²

1973

Plant Physiol.

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Cotyledons of Phaseolus aureus contain protein-bound glu. cosamine which is metabolized during germination. The glucosamine is present in storage glycoproteins, and these are concurrently metabolized along with the glucosamine. These glycoproteins are associated with protein bodies. Characterization of the glucosamine-containing storage proteins showed them to be identical with vicilin and legumin, the major storage proteins of the Leguminosae. Extraction of Proteins. The cotyledons were homogenized either in 0.4 M NaCl or in 20 mm sodium borate, pH 9.0, using 2 ml of medium per g of tissue. The homogenate was centrifuged at 30,000g for 20 min, and the supernatant was collected. The pellet was extracted three times with the same medium, and the extracts were combined with the initial supernatant.Quantitative Determination of Protein. Proteins were usually assayed according to the procedure of Lowry et al. (13) using bovine serum albumin as a standard. The protein content of peak III glycoprotein was also determined with ninhydrin (27) after acid hydrolysis of the protein.Quantitative determination of hexosamine. The samples to be assayed for protein-bound hexosamine were precipitated with 7.5% trichloroacetic acid, washed with 5% trichloroacetic acid, alcohol, and ether. The dried samples were resuspended in 1 N HCI. Samples to be assayed for total hexosamine were homogenized in 1 N HCl. Hydrolysis was carried out at 90 C for 24 hr. Under these conditions there was no detectable loss of hexosamine. Hexosamines were separated from interfering chromogens by passage through a small Dowex 50 cation exchange column according to the method of Boas (4). This step is absolutely necessary to obtain reproducible results. Eluates from the columns were dried by evaporation under vacuum at 85 C, and the residue was dissolved in water. The hexosamine content was determined by the method of Elson-Morgan as modified by Johnson (11

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confidence: 76%

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confidence: 92%

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confidence: 99%

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Isolation and Characterization of Glucosamine-containing Storage Glycoproteins from the Cotyledons of Phaseolus aureus

Ericson¹,

Chrispeels²

1973

Plant Physiol.

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“…Alternatively, although the relative mobilities of the LB subunits in urea gels at alkaline pH appear to be consistent with our definition of them as the xmsubunits, it is possible that there is some aggregation of legumin subunits in 8M urea at alkaline pH (cf. Grant and Lawrence, 1964) and that the inclusion of Nonidet and higher concentrations (9 5M) of urea reduces or eliminate aggregation; hence LB subunits and ocmsubunits may not be strictly comparable. The LC system refers to slight differences in the subunit molecular weight of the major 40 x 10 mol.…”

Section: Dxscussiomentioning

confidence: 99%

Genetic variability in the structure of the α-subunits of legumin from Pisum—a two-dimensional gel electrophoresis study

Casey

1979

Heredity

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SUMMARYVariant forms of legumin (one of the major storage globulins of Pisum seeds) were purified by immunoaffinity chromatography from a range of Pisum types, includingP.fulvum, and primitiveforms, modern cultivars and advanced breeding selections of P. sativuin. The purified variants, all of which had sedimentation coefficients of about 12S and leucine, threonine and glycine as N-terminal amino acids, showed subunit heterogeneity on polyacrylamide gels containing sodium dodecyl sulphate and on two-dimensional isofocusing/electrophoresis. Comparison of legumins from different Pisum types showed variability, both in the net surface charge of the whole molecule and in the nature of subunit heterogeneity on two-dimensional gels. The usefulness of two-dimensional gels in the genetic analysis of pea seed storage protein structure was stressed and the role of potential artefacts in such analyses assessed.

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“…1 Considerable difficulty has been encountered in separating completely legumin from vicilin by ammonium sulfate precipitation (11,19,28). Chromatography on diethylaminoethyl cellulose has also been used (15,27). In our hands, this yielded legumin contaminated with small molecular weight proteins.…”

Section: Isolation and Characterisation Of Legumin And Vicilinmentioning

confidence: 99%

Legumin Synthesis in Developing Cotyledons of Vicia faba L.

1971

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The synthesis of legumin in developing cotyledons of Vicia faba L. has been examined as a potential system for approaching the problem of differential gene expression. The pattern of legumin synthesis was determined during the growth of the cotyledon by microcomplement fixation which provided a sensitive and specific assay for legumin in the presence of vicilin. Legumin was detected even in young cotyledons. However, when the cotyledons were about 10 millimeters long, and cell division was essentially complete, there was a sharp increase in the rate of legumin accumulation.Developing seeds of Leguminosae offer an attractive system for studying the problem of differential gene expression. These seeds accumulate large amounts of globulins which consist of two high molecular weight proteins, legumin and vicilin, as defined by analytical centrifugation (11,12). Varner and Schidlovsky (29) showed globulins to be localized in protein bodies within pea cotyledons. A major attempt to use developing seeds of Leguminosae for studies of differential gene expression was first made by . They used immature seeds of Pisum sativum for analyzing the regulation of transcription and translation of chromatin preparations from cotyledon cells. More recently, Boulter and associates (2,3,9,10,25,32) have initiated extensive studies of cotyledon development in V.faba. In this paper we wish to report our analyses of legumin synthesis in relation to the growth of the cotyledon of V. faba. MATERIAIS AND METHODS Plant Material. Seeds of Vicia faba L. (cultivar unknown)were grown in soil in the greenhouse or garden. When pods were harvested, any abnormally shaped material was excluded. In a pod there were usually three or four seeds of equal size which were used and one or two smaller seeds which were

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Effects of sodium dodecyl sulfate and other dissociating reagents on the globulins of peas

Cited by 46 publications

References 27 publications

Isolation and Characterization of Glucosamine-containing Storage Glycoproteins from the Cotyledons of Phaseolus aureus

Isolation and Characterization of Glucosamine-containing Storage Glycoproteins from the Cotyledons of Phaseolus aureus

Genetic variability in the structure of the α-subunits of legumin from Pisum—a two-dimensional gel electrophoresis study

Legumin Synthesis in Developing Cotyledons of Vicia faba L.

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