An electrophoretic analysis of the seed proteins from Pinus monticola and eight other species of pine

1988 ◽  
Vol 66 (9) ◽  
pp. 1808-1812 ◽  
Author(s):  
David J. Gifford
Keyword(s):  
Molecular Mass ◽  
Storage Proteins ◽  
Ricinus Communis ◽  
Sodium Dodecyl ◽  
Reduced Form ◽  
Major Protein ◽  
Polyacrylamide Gels ◽  
Pinus Monticola ◽  
Embryonic Axes ◽  
Pinus Species

The major storage proteins of mature Pinus monticola Dougl. seed megagametophytes are crystalloids and are only completely soluble in buffer solutions if sodium dodecyl sulphate is present. These insoluble proteins-constitute 50% of the storage reserve in the seed. Crystalloid proteins are found in embryonic axes also and are a major protein reserve in mature seeds from all other Pinus species examined. These proteins in their nonreduced form have molecular masses of 51 – 55 kilodaltons (kDa) and in reduced form migrate on polyacrylamide gels as two distinct groups of proteins, one in the molecular mass range 31 – 34.5 kDa and the other in the 21.5 – 22.5 kDa range. The pine crystalloid proteins are not glycosylated and have similar solubility, structural, and size characteristics to crystalloid proteins found in other seed types, such as Ricinus communis and Cucurbita species. However, they are immunologically different from the crystalloid proteins of R. communis. In general, the soluble protein gel profiles vary considerably among the species. Some similarities do exist; in particular, a group of proteins in the 27 – 29.5 kDa molecular mass regions of polyacrylamide gels is common to all Pinus species examined. The soluble proteins are not glycosylated.

A metalloproteinase extracellularly released byCrithidia deanei

2003 ◽  
Vol 49 (10) ◽  
pp. 625-632 ◽  
Author(s):  
Claudia Masini d'Avila-Levy ◽  
Rodrigo F Souza ◽  
Rosana C Gomes ◽  
Alane B Vermelho ◽  
Marta H Branquinha
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Residual Activity ◽  
Major Protein ◽  
Motile Cells ◽  
Sds Page ◽  
Triton X

Actively motile cells from a cured strain of Crithidia deanei released proteins in phosphate buffer (pH 7.4). The molecular mass of the released polypeptides, which included some proteinases, ranged from 19 to 116 kDa. One of the major protein bands was purified to homogeneity by a combination of anion-exchange and gel filtration chromatographs. The apparent molecular mass of this protein was estimated to be 62 kDa by sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE). The incorporation of gelatin into SDS–PAGE showed that the purified protein presented proteolytic activity in a position corresponding to a molecular mass of 60 kDa. The enzyme was optimally active at 37 °C and pH 6.0 and showed 25% of residual activity at 28 °C for 30 min. The proteinase was inhibited by 1,10-phenanthroline and EDTA, showing that it belonged to the metalloproteinase class. A polyclonal antibody to the leishmanial gp63 reacted strongly with the released C. deanei protease. After Triton X-114 extraction, an enzyme similar to the purified metalloproteinase was detected in aqueous and detergent-rich phases. The detection of an extracellular metalloproteinase produced by C. deanei and some other Crithidia species suggests a potential role of this released enzyme in substrate degradation that may be relevant to the survival of trypanosomatids in the host.Key words: endosymbiont, trypanosomatid, extracellular, proteinase.

A comparative study of the insoluble storage proteins and the lectins of seeds of the Euphorbiaceae

1984 ◽  
Vol 62 (8) ◽  
pp. 1671-1677 ◽  
Author(s):  
Louise Lalonde ◽  
David W. Fountain ◽  
Allison Kermode ◽  
Francis B. Ouellette ◽  
Kelly Scott ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Castor Bean ◽  
Storage Proteins ◽  
Reduced Form ◽  
Polyacrylamide Gels ◽  
Salt Solutions ◽  
Sodium Dodecylsulphate ◽  
Molecular Weight Range ◽  
Jatropha Gossypifolia ◽  
Major Storage Protein

The major storage protein within seeds of the Euphorbiaceae is the 11S crystalloid, which is only completely soluble in buffer or salt solutions if sodium dodecylsulphate or urea is present. Prior to this study, only the storage proteins of the castor bean had been characterized. The nonreduced crystalloid protein complex in all species tested has a molecular weight of 50 000 – 55 000, and in reduced form the proteins migrate on polyacrylamide gels as two distinct groups of polypeptides, one in the molecular weight range 20 000 – 25 000 and the other in the 29 000 – 35 000 range. In this respect the proteins have the general characteristics of those of castor bean, but only the proteins of Jatropha gossypifolia show striking similarities. Within any one genus, the storage proteins appear to be more or less identical (e.g., Manihot spp.) or show distinct differences (e.g., Euphorbia spp.). The soluble lectin proteins of J. gossypifolia have very similar haemagglutination properties to those of castor bean lectins, and the glycoproteins of both species separate similarly on polyacrylamide gels. Few other species contain glycoproteins or lectins that can cause agglutination.

Efficacy and compatibility with mass spectrometry of methods for elution of proteins from sodium dodecyl sulfate–polyacrylamide gels and polyvinyldifluoride membranes

2004 ◽  
Vol 330 (1) ◽  
pp. 87-97 ◽  
Author(s):  
Charlotte Sværke Jørgensen ◽  
Mitchell Jagd ◽  
Birgitte Kjær Sørensen ◽  
Jim McGuire ◽  
Vibeke Barkholt ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gels ◽  
Dodecyl Sulfate

SELECTION OF COLUMN AND OPERATING CONDITIONS FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PROTEINS IN CANADIAN WHEAT

1985 ◽  
Vol 65 (2) ◽  
pp. 285-298 ◽  
Author(s):  
J. E. KRUGER ◽  
B. A. MARCHYLO
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Storage Proteins ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Operating Conditions ◽  
Protein Patterns ◽  
Optimal Separation ◽  
Rp Hplc

Chromatographic conditions were optimized and three commercially available columns were evaluated for separation of alcohol-soluble storage proteins of Neepawa wheat using reversed-phase high-performance liquid chromatography (RP-HPLC). Optimal separation was achieved using an extracting solution of 50% 1-propanol, 1% acetic acid, and 4% dithiothreitol and an HPLC elution time of 105 min at a flow rate of 1.0 mL/min. HPLC columns evaluated (SynChropak RP-P, Ultrapore RPSC and Aquapore RP-300) varied in selectivity and resolution. The column providing the greatest versatility was Aquapore RP-300 available in cartridge form. Sodium dodecyl sulfate gradient-gel electrophoresis analysis of protein peaks resolved by RP-HPLC indicated that many of the eluted peaks contained more than one protein species. Chromatographic protein patterns obtained for Neepawa wheat grown at different locations and in different years were qualitatively the same.Key words: Protein, high-performance liquid chromatography, wheat

Calcium-binding proteins in a vorticellid contractile organelle

1975 ◽  
Vol 19 (1) ◽  
pp. 203-213
Author(s):  
W.B. Amos ◽  
L.M. Routledge ◽  
F.F. Yew
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Calcium Binding ◽  
Binding Proteins ◽  
Sodium Dodecyl ◽  
Aromatic Amino Acids ◽  
Calcium Binding Proteins ◽  
Polyacrylamide Gels ◽  
Protein Species ◽  
Low Concentrations

The proteins of the contractile spasmoneme of Zoothamnium have been examined for comparison with other motile systems. Though capable of calcium-induced contraction, glycerinated preparations of the spasmoneme contain neither actin nor tubulin at levels that can be detected in polyacrylamide gels. Sixty per cent of the protein in sodium dodecyl sulphate gels migrates in a band at a molecular weight of approximately 20,000, consisting largely of 2 similar protein species which are here given the name of spasmins. The amino acid composition of 2 spasmin fractions has been determined by a fluorimetric method. They are rich in Asx, Glx and serine, but have few aromatic amino acids and no cystine or methionine. In calcium-buffered polyacrylamide gels, it was observed that a reduction in the electrophoretic mobility of the spasmins was induced specifically by calcium (but not magnesium) at the same low concentrations as induce contraction. This indicates that the spasmins are calcium-binding proteins which may be involved directly in the calcium-induced contraction of the spasmoneme.

scholarly journals Reciprocal effect of parental lines on the physiological potential and seed composition of corn hybrid seeds

2017 ◽  
Vol 27 (3) ◽  
pp. 206-216 ◽  
Author(s):  
Juliana F. Santos ◽  
Lynnette M.A. Dirk ◽  
A. Bruce Downie ◽  
Mauricio F.G. Sanches ◽  
Roberval D. Vieira
Keyword(s):  
Storage Proteins ◽  
Soluble Sugar ◽  
Soluble Sugars ◽  
Major Protein ◽  
Seed Composition ◽  
Seed Vigour ◽  
Reciprocal Crosses ◽  
Hybrid Seeds ◽  
Parental Lines ◽  
Corn Hybrid

AbstractObtaining corn hybrid seeds (Zea mays L.) with high vigour depends on the parental lines and the direction of the cross, and this relates to seed desiccation tolerance and composition. This research studied reciprocal crosses between pairs of proprietary, elite parent lines (L1 and L5; L2 and L4) producing hybrid seeds with different qualities attempting to correlate vigour with seed composition, focusing on storage proteins, starch and soluble sugar amounts. Four corn hybrid seed lots produced from reciprocal crosses were compared (HS 15 with HS 51, and HS 24 with HS 42) by assessing germination, vigour, and seedling emergence in the field. Seed composition was assessed in mature, dehydrated seeds. Proteins were extracted, quantified, and analysed by electrophoresis and densitometry. Starch amounts were assessed using a kit and soluble sugars were determined using high performance liquid chromatography with pulsed electrochemical detection. The L1 and L2 lineages, used as female parents, provided seeds with lower vigour; however, the quantification of major protein bands, and sucrose, raffinose and stachyose were similar between seed lot pairs. While both total seed protein and starch varied between reciprocal hybrids for one of the two sets of crosses, the amounts of neither correlated with seed vigour. Interestingly, hybrids with low seed vigour (HS 15, HS 24) accumulated greater amounts of fructose relative to their reciprocal; correlation analysis confirmed these results. We demonstrate different effects on seed vigour dependent on the maternal parent in reciprocal crosses producing hybrid corn seeds. We also show that vigour is negatively correlated with seed reducing sugar contents.

Abscisic acid and low temperature induced polypeptide changes in alfalfa (Medicago sativa) cell suspension cultures

1986 ◽  
Vol 64 (11) ◽  
pp. 2758-2763 ◽  
Author(s):  
Albert J. Robertson ◽  
Lawrence V. Gusta
Keyword(s):  
Abscisic Acid ◽  
Low Temperature ◽  
Medicago Sativa ◽  
Cell Suspension ◽  
Sodium Dodecyl ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Total Cell ◽  
Cell Protein ◽  
Major Protein

Changes in extracellular, cellular, and subcellular proteins during abscisic acid and low temperature induced cold hardening of alfalfa (Medicago sativa L. cv. Wisconsin 22C) cell suspension cultures were investigated by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Extracellular proteins from 4- to 6-day-old ABA and low temperature grown alfalfa cells showed decreased electrophoretic mobilities, lacked a 190-kDa glycoprotein, and had reduced amounts of four other polypeptides. In total cell protein analyses, a 42-kDa protein was enriched in both ABA and low temperature treated alfalfa cells. Several proteins increased or induced by exogenous ABA treatment were identified in the extracellular (12.5 and 13 to 15 kDa), total cell and cell wall (24 kDa), and soluble (20, 37, and 41 kDa) fractions. However, no major protein changes were resolved by one-dimensional electrophoretic analyses of crude membrane proteins.

Purification and Partial Amino Acid Sequence of Pentocin 31-1, an Anti-Listeria Bacteriocin Produced by Lactobacillus pentosus 31-1

2009 ◽  
Vol 72 (12) ◽  
pp. 2524-2529 ◽  
Author(s):  
JINLAN ZHANG ◽  
GUORONG LIU ◽  
NAN SHANG ◽  
WANPENG CHENG ◽  
SHANGWU CHEN ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Specific Activity ◽  
Consensus Sequence ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Mass Spectrometry Analysis ◽  
Gel Chromatography ◽  
Original Activity ◽  
Lactobacillus Pentosus ◽  
Spectrometry Analysis

Pentocin 31-1, an anti-Listeria bacteriocin produced by Lactobacillus pentosus 31-1 from the traditional Chinese fermented Xuan-Wei ham, was successfully purified by the pH-mediated cell adsorption-desorption method and then purified by gel chromatography with Sephadex G-10. The purification resulted in a 1,381.9-fold increase in specific activity with a yield of 76.8% of the original activity. Using Tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), the molecular mass of the purified peptide was found to be between 3,500 and 6,400 Da, and bacteriocin activity was confirmed by overlayer techniques. When subjected to mass spectrometry analysis, the protein was highly pure and its molecular mass was 5,592.225 Da. The partial N-terminal sequence of pentocin 31-1 was the following: NH2-VIADYGNGVRXATLL. Compared with the sequence of other bacteriocins, pentocin 31-1 has the consensus sequence YGNGV in its N-terminal region, and therefore it belongs to the class IIa of bacteriocins.

Major polypeptides associated with differentiation in psychrophilic fungi

1987 ◽  
Vol 65 (2) ◽  
pp. 233-241 ◽  
Author(s):  
W. J. Newsted ◽  
N. P. A. Huner
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Molecular Mass ◽  
Relative Proportion ◽  
Sodium Dodecyl ◽  
Isoelectric Points ◽  
Dodecyl Sulfate ◽  
Typhula Incarnata ◽  
Culturing Conditions ◽  
Sclerotial Development ◽  
Polypeptide Profile

Major polypeptides were observed upon one-dimensional sodium dodecyl sulfate gel electrophoresis of sclerotial extracts of the following psychrophiles: Myriosclerotinia borealis, Coprinus psychromorbidus, Typhula idahoensis, and Typhula incarnata. In general, the number, molecular mass, and relative proportion of these major sclerotial polypeptides varied considerably from species to species. Furthermore, in the case of M. borealis, the major sclerotial polypeptide did not appear to be an artifact of culturing conditions since a major polypeptide of similar molecular mass was also present in sclerotia of M. borealis collected from the field. Generally, the major sclerotial polypeptides were visible in the sclerotial initials but were not apparent in the vegetative hyphae. Thus, these major sclerotial polypeptides appear to be expressed as a function of sclerotial development. Electrophoresis of protein extracts of T. idahoensis and T. incarnata initially solubilized either in sodium dodecyl sulfate or urea sample buffer indicated that the type of denaturant initially used had a profound influence on the relative proportions of the major polypeptides and the overall polypeptide profile. Isoelectric focusing of sclerotial extracts indicated that the isoelectric points of the major sclerotial polypeptides of M. borealis ranged from 6.2 to 6.7, whereas the values of the major sclerotial polypeptides of the other three species were basic and ranged from 7.0 to 7.7.

Sign in / Sign up

Export Citation Format

Share Document