cDNA and protein sequence of a major pea seed albumin (PA 2 : Mr?26 000)

Higgins, T. J. V.; Beach, Larry R.; Spencer, Donald; Chandler, Peter M.; Randall, P. J.; Blagrove, R.J.; Kortt, Alexander A.; Guthrie, Robin E.

doi:10.1007/bf00016432

Cited by 54 publications

(71 citation statements)

References 30 publications

(34 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The gene encodes a polypeptide that has the signal peptide and first three amino acids of PHA-L, and the coding sequence of a pea seed albumin (12). Two additional amino acids were introduced when the translational fusion was made.…”

Section: Resultsmentioning

confidence: 99%

The Signal Peptide of a Vacuolar Protein Is Necessary and Sufficient for the Efficient Secretion of a Cytosolic Protein

Hunt¹,

Chrispeels²

1991

Plant Physiol.

View full text Add to dashboard Cite

A cytosolic pea (Pisum sativum) seed albumin (ALB) and a chimeric protein (PHALB) consisting of the signal peptide and first three amino acids of phytohemagglutinin (PHA) and the amino acid sequence of ALB were expressed in parallel suspension cultures of tobacco (Nicotiana tabacum) cells and their intracellular fates examined. PHALB was efficiently secreted by the cells whereas ALB remained intracellular. These experiments show that the information contained in the signal peptide of a vacuolar protein is both necessary and sufficient for efficient secretion, and define secretion as a default or bulk-flow pathway. Entry into the secretary pathway was accompanied by glycosylation and the efficient conversion of the high mannose glycans into complex glycans indicating that transported glycoproteins do not need specific recognition domains for the modifying enzymes in the Golgi. Tunicamycin depressed the accumulation of the unglycosylated polypeptide in the culture medium much less than the accumulation of other glycoproteins. We interpret this as evidence that glycans on proteins that are not normally glycosylated do not have the same function of stabilizing and protecting the polypeptide as on natural glycoproteins.The secretary system of plant cells delivers proteins to the vacuole, the tonoplast, the plasma membrane, and the cell wall/extracellular space. In addition, proteins that enter the secretary system may be retained in the endoplasmic reticulum or in various compartments of the Golgi complex. The first step common to the transport of all these proteins is translocation across the ER membrane (25). Once

show abstract

Section: Resultsmentioning

confidence: 99%

The Signal Peptide of a Vacuolar Protein Is Necessary and Sufficient for the Efficient Secretion of a Cytosolic Protein

Hunt¹,

Chrispeels²

1991

Plant Physiol.

View full text Add to dashboard Cite

show abstract

“…Numerous research attempts have been made to isolate, separate and identify pea albumins and determine their allergenic potential [Croy et al 1984, Higgins et al 1987. The cited authors isolated, separated and identifi ed two main pea albumin fractions (Fig.…”

Section: Resultsmentioning

confidence: 99%

Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds

Dziuba

Szerszunowicz

Nałęcz

et al. 2014

Acta Sci Pol Technol Aliment

View full text Add to dashboard Cite

Background. Proteomic analysis is emerging as a highly useful tool in food research, including studies of food allergies. Two-dimensional gel electrophoresis involving isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis is the most effective method of separating hundreds or even thousands of proteins. In this study, albumin and globulin fractions of pea seeds cv. Ramrod were subjected to proteomic analysis. Selected potentially alergenic proteins were identifi ed based on their molecular weights and isoelectric points. Material and methods. Pea seeds (Pisum sativum L.) cv. Ramrod harvested over a period of two years (Plant Breeding Station in Piaski-Szelejewo) were used in the experiment. The isolated albumins, globulins and legumin and vicilin fractions of globulins were separated by two-dimensional gel electrophoresis. Proteomic images were analysed in the ImageMaster 2D Platinum program with the use of algorithms from the Melanie application. The relative content, isoelectric points and molecular weights were computed for all identifi ed proteins. Electrophoregrams were analysed by matching spot positions from three independent replications. Results. The proteomes of albumins, globulins and legumin and vicilin fractions of globulins produced up to several hundred spots (proteins). Spots most characteristic of a given fraction were identifi ed by computer analysis and spot matching. The albumin proteome accumulated spots of relatively high intensity over a broad range of pI values of ~4.2-8.1 in 3 molecular weight (MW) ranges: I -high molecular-weight albumins with MW of ~50-110 kDa, II -average molecular-weight albumins with MW of ~20-35 kDa, and III -low molecular-weight albumins with MW of ~13-17 kDa. 2D gel electrophoregrams revealed the presence of 81 characteristic spots, including 24 characteristic of legumin and 14 -of vicilin. Conclusions. Two-dimensional gel electrophoresis proved to be a useful tool for identifying pea proteins. Patterns of spots with similar isoelectric points and different molecular weights or spots with different isoelectric points and similar molecular weights play an important role in proteome analysis. The regions characteristic of albumin, globulin and legumin and vicilin fractions of globulin with typical MW and pI values were identifi ed as the results of performed 2D electrophoretic separations of pea proteins. 2D gel electrophoresis of albumins and the vicilin fraction of globulins revealed the presence of 4 and 2 spots, respectively, representing potentially allergenic proteins. They probably corresponded to vicilin fragments synthesized during post-translational modifi cation of the analysed protein.

show abstract

“…A lack of PA2 was accompanied by an increase in the levels of Ala, Phe, Cys, Arg, and Gln. Because accumulation of PA2 contributes approximately 11% to the sulfur-amino acids in pea seeds (Higgins et al, 1987), the increase in free Cys could be the direct consequence of decreased PA2 synthesis. The increase in the N-rich amino acids Arg and Gln and the increase in the Gln to Glu ratio indicate that lack of PA2 leads to a higher N status in the seeds.…”

Section: Discussionmentioning

confidence: 99%

“…Because the major part of PA2 accumulation occurs from 20 d after flowering (DAF) onwards (Chandler et al, 1984;Higgins et al, 1987;Fig. 5, tracks a-c), metabolite levels were measured at different stages of seed development.…”

Section: Metabolite Profiling Of Pa2 Variant Linesmentioning

confidence: 99%

See 1 more Smart Citation

Combined Metabolomic and Genetic Approaches Reveal a Link between the Polyamine Pathway and Albumin 2 in Developing Pea Seeds

et al. 2007

View full text Add to dashboard Cite

Several legume seed proteins that are potentially allergenic, poorly digested by farm animals, and/or have undesirable functional properties, have been described. One of these is the albumin protein in pea (Pisum sativum) called PA2. A naturally occurring mutant line that lacks PA2 has been exploited in studies to determine the biological function of this nonstorage protein in seed development. The mutant, which has a small seed, a tall plant phenotype, and lacks most of the PA2-encoding genes, has been crossed with a standard cultivar, 'Birte,' which contains PA2 to give rise to a recombinant inbred (RI) population. An F 3 line carrying the mutation and having a short plant phenotype has been used to generate backcross (BC) lines with 'Birte.' Despite having a lower albumin content, seeds from the mutant parent and RI lines lacking PA2 have an equivalent or higher seed nitrogen content. Metabolite profiling of seeds revealed major differences in amino acid composition and polyamine content in the two parent lines. This was investigated further in BC lines, where the effects of differences in seed size and plant height between the two parents were eliminated. Here, differences in polyamine synthesis were maintained as was a difference in total seed protein between the BC line lacking PA2 and 'Birte.' Analysis of enzyme activities in the pathways of polyamine synthesis revealed that the differences in spermidine content were attributable to changes in the overall activities of spermidine synthase and arginine decarboxylase. Although the genes encoding spermidine synthase and PA2 both localized to the pea linkage group I, the two loci were shown not to be closely linked and to have recombined in the BC lines. A distinct locus on linkage group III contains a gene that is related to PA2 but expressed predominantly in flowers. The results provide evidence for a role of PA2 in regulating polyamine metabolism, which has important functions in development, metabolism, and stress responses in plants.

show abstract

cDNA and protein sequence of a major pea seed albumin (PA 2 : Mr?26 000)

Cited by 54 publications

References 30 publications

The Signal Peptide of a Vacuolar Protein Is Necessary and Sufficient for the Efficient Secretion of a Cytosolic Protein

The Signal Peptide of a Vacuolar Protein Is Necessary and Sufficient for the Efficient Secretion of a Cytosolic Protein

Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds

Combined Metabolomic and Genetic Approaches Reveal a Link between the Polyamine Pathway and Albumin 2 in Developing Pea Seeds

Contact Info

Product

Resources

About