Cold Acclimation and Alterations in Dehydrin-like and Bark Storage Proteins in the Leaves of Sibling Deciduous and Evergreen Peach

Seasonal changes in cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach [Prunus persica (L.) Batsch]. Freezing tolerance [defined as the subzero temperature at which 50% injury occurred (LT50)] was assessed using electrolyte leakage. Proteins were separated by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. Electroblots were probed with anti-dehydrin and anti-19-kD peach bark storage protein (BSP) antibodies. Leaf LT50 decreased successively from -5.8 °C on 18 Aug. to -10.3 °C in the evergreen genotype and from -7.0 °C to -15.0 °C in the deciduous genotype by 14 Oct. Protein profiles and immunoblots indicated the accumulation of a 60- and 30-kD protein during cold acclimation in the leaves of deciduous trees; however, levels of these proteins did not change significantly in the evergreen trees. Immunoblots indicate that the 60-kD protein is a dehydrin-like protein. Gel-electrophoresis and immunoblots also indicated that the 19-kD BSP progressively disappeared from summer through fall in leaves of deciduous peach but accumulated to a high level in bark tissues. A similar inverse relationship was not evident in evergreen peach.

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Diversity of seed storage proteins in common wheat (Triticum aestivum L.)

Plant Genetic Resources ◽

10.1017/s1479262111000554 ◽

2011 ◽

Vol 9 (2) ◽

pp. 256-259

Author(s):

Zuzana Šramková ◽

Edita Gregová ◽

Svetlana Šliková ◽

Ernest Šturdík

Keyword(s):

Triticum Aestivum ◽

Common Wheat ◽

Gel Electrophoresis ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Seed Storage ◽

Seed Storage Proteins ◽

Triticum Aestivum L ◽

Polyacrylamide Gel

The objective of our study was to determine the composition of high-molecular weight-glutenin subunits (HMW-GS) in 120 cultivars of common wheat (Triticum aestivum L.). Fourteen alleles and 34 allelic compositions were detected using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The most frequent HMW-GS alleles at the Glu-A1, Glu-B1 and Glu-D1 loci were null (57.1%), 7+9 (43.3%) and 5+10 (61.9%), respectively. However, low-frequency HMW-GS alleles were also observed, such as 13+16, 20, 21, 7 and 18, encoded by the Glu-B1 locus, and 4+12, encoded by the Glu-D1 locus. The wheat–rye 1BL.1RS translocation was identified in 25 cultivars, using acid polyacrylamide gel electrophoresis. The Glu-score varied greatly, and some lines reached the maximum value of 10.

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COLD ACCLIMATION IN THE LEAVES OF SIBLING DECIDUOUS AND EVERGREEN PEACH: ALTERATIONS IN DEHYDRIN AND BARK STORAGE PROTEINS

HortScience ◽

10.21273/hortsci.30.2.190f ◽

1995 ◽

Vol 30 (2) ◽

pp. 190f-191

Author(s):

Rajeev Arora ◽

Michael Wisniewski ◽

Lisa J. Rowland

Keyword(s):

Cold Acclimation ◽

Cold Tolerance ◽

Prunus Persica ◽

Storage Proteins ◽

Seasonal Pattern ◽

Bark Tissue ◽

Bark Storage Proteins ◽

Source Sink ◽

Favorable Conditions ◽

Bark Storage Protein

Seasonal pattern of cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach (Prunus persica). In contrast to deciduous peach that undergoes endodormancy in fall, evergreen peach does not (leaves are retained and shoot tips elongate under favorable conditions) (Arora et al., Plant Physiol. 99:1562-1568). Cold tolerance (LT50) was assessed using electrolyte leakage method. Proteins were separated by SDS-PAGE. Electroblots were probed with anti-dehydrin (Dr. T. Close) and anti-19 kD, peach bark storage protein (BSP) antibodies. LT50 of leaves successively increased from about -7C (18 Aug.) to -15C and -11.5C (23 Oct.) in deciduous and evergreen genotypes, respectively. The most apparent change in the protein profiles was the accumulation of a 60-kD protein during cold acclimation in the leaves of deciduous trees; however, it did not change significantly in evergreen peach. Immunoblots indicate that 60-kD protein is a dehydrin protein. PAGE and immunoblots indicated that 19-kD BSP disappeared progressively during summer through fall in the leaves of deciduous peach, but accumulated to large amounts in bark tissues. Similar inverse relationship for its accumulation in leaf vs. bark tissue was not evident in evergreen peach. Results indicate that BSP expression may be regulated by altered source/sink relationship.

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c-Type Cytochromes in Pelobacter carbinolicus

Applied and Environmental Microbiology ◽

10.1128/aem.01128-06 ◽

2006 ◽

Vol 72 (11) ◽

pp. 6980-6985 ◽

Cited By ~ 20

Author(s):

Shelley A. Haveman ◽

Dawn E. Holmes ◽

Yan-Huai R. Ding ◽

Joy E. Ward ◽

Raymond J. DiDonato ◽

...

Keyword(s):

Mass Spectrometry ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Geobacter Sulfurreducens ◽

Open Reading Frames ◽

Genetic Studies ◽

Close Relatives ◽

High Level ◽

Reading Frames

ABSTRACT Previous studies failed to detect c-type cytochromes in Pelobacter species despite the fact that other close relatives in the Geobacteraceae, such as Geobacter and Desulfuromonas species, have abundant c-type cytochromes. Analysis of the recently completed genome sequence of Pelobacter carbinolicus revealed 14 open reading frames that could encode c-type cytochromes. Transcripts for all but one of these open reading frames were detected in acetoin-fermenting and/or Fe(III)-reducing cells. Three putative c-type cytochrome genes were expressed specifically during Fe(III) reduction, suggesting that the encoded proteins may participate in electron transfer to Fe(III). One of these proteins was a periplasmic triheme cytochrome with a high level of similarity to PpcA, which has a role in Fe(III) reduction in Geobacter sulfurreducens. Genes for heme biosynthesis and system II cytochrome c biogenesis were identified in the genome and shown to be expressed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels of protein extracted from acetoin-fermenting P. carbinolicus cells contained three heme-staining bands which were confirmed by mass spectrometry to be among the 14 predicted c-type cytochromes. The number of cytochrome genes, the predicted amount of heme c per protein, and the ratio of heme-stained protein to total protein were much smaller in P. carbinolicus than in G. sulfurreducens. Furthermore, many of the c-type cytochromes that genetic studies have indicated are required for optimal Fe(III) reduction in G. sulfurreducens were not present in the P. carbinolicus genome. These results suggest that further evaluation of the functions of c-type cytochromes in the Geobacteraceae is warranted.

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Plasma protein concentrations of the young and adult Amazona brasiliensis parrots

Veterinarski glasnik ◽

10.2298/vetgl170117008s ◽

2017 ◽

Vol 71 (1) ◽

pp. 44-51

Author(s):

Elizabeth Schmidt ◽

Patricia Serafini ◽

Elenise Sipinski ◽

Antonio Paulillo

Keyword(s):

Plasma Protein ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Clinical Pathology ◽

Specific Protein ◽

Major Protein ◽

Plasma Protein Concentration ◽

Protein Gel ◽

Sds Page

Introduction. The Red-tailed Amazon parrot (Amazona brasiliensis) is an endangered species of the Psittacine family, and for which various data are important for a comprehensive preservation plan. Data about plasma protein gel electrophoresis of Amazon parrot blood are scarce. The purpose of this study was to determine plasma protein concentrations and concentrations of major protein bands in blood of young and adult Red-tailed Amazon parrot (Amazona brasiliensis). Materials and Methods. Blood samples from eight young and eight adult healthy free-living parrots were obtained. Plasma protein concentration and fractions were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Mann-Whitney U test was used to compare variables. Results and Conclusions. Six major protein bands with the following molecular weights were identified by SDS-PAGE: 170 kDa, 117 kDa, 85 kDa (putative ovotransferrin), 60 kDa, 45 kDa and 23 kDa. Adult parrots had significantly higher concentrations of total proteins, albumin and other proteins with similar mobility (around 60 kDa). Young birds had significantly higher levels of 23kDa proteins. The concentration of putative ovotransferrin (85 kDa) was not different between young and adult parrots. Plasma protein gel electrophoresis patterns in Red-tailed Amazon parrots are similar between young and adult animals, but specific protein bands differ in their absolute concentrations. This finding should be taken into consideration when clinical pathology data are analysed.

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Genetic Effects on the Proportions of Conglutin γ and a Subunit of Conglutin β in the Storage Proteins of Lupinus angustifolius L. Seeds

Functional Plant Biology ◽

10.1071/pp9810277 ◽

1981 ◽

Vol 8 (3) ◽

pp. 277 ◽

Cited By ~ 3

Author(s):

RN Oram ◽

JM Gillespie ◽

RJ Blagrove

Keyword(s):

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Seed Storage ◽

Seed Storage Proteins ◽

Lupinus Angustifolius ◽

Β Subunit ◽

Partial Dominance ◽

High Level ◽

Conglutin Γ

The genetic variability in the proportions of conglutin β and γ components of lupin seed storage proteins was explored in F1, F2 and F3 families generated from a cross between two genotypes of Lupinus angustifolius cv. Uniharvest and a white-seeded selection from CPI 47644. The levels of conglutin γ differed by a factor of two in these parents, but varied continuously in the F2 and F3 families, indicating that the parental difference is due to several genes. These parents also differed in the level of a major conglutin β subunit, with a molecular weight of 30 000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the storage globulins from individual F1, F2 and F3 seeds showed that the high level in cv. Uniharvest is controlled by an incompletely dominant allele, Bsc, at one locus, whereas the low level in CPI 47644 (white-seeded) is due to homozygosity for the recessive allele, bsc. The partial dominance of the Uniharvest gene suggests that the locus has a regulatory or processing function, so it has been designated beta subunit controller. F3 families differed significantly in the level of the conglutin β subunit in Bsc/- seeds, indicating that other loci with smaller effects also influence the proportion of this subunit. The proportion of the β subunit in any seed depends largely on its own genotype, and not on the genotype of the maternal plant. The proportion of conglutin γ in the storage proteins varied inversely with the proportion of the β subunit. The implications of these results for the genetic improvement of the biological value of lupin protein are discussed.

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Proteome analysis of two soft-grained wheats of different processing quality: cultivar-specific proteins

Australian Journal of Agricultural Research ◽

10.1071/ar04114 ◽

2005 ◽

Vol 56 (2) ◽

pp. 145 ◽

Cited By ~ 7

Author(s):

D. J. Skylas ◽

S. J. Cordwell ◽

G. Craft ◽

B. McInerney ◽

M. J. Wu ◽

...

Keyword(s):

Gel Electrophoresis ◽

Starch Granule ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Proteome Analysis ◽

Specific Protein ◽

Processing Quality ◽

Ph Gradients ◽

Specific Proteins

Proteome analysis was conducted on the grain of 2 closely related soft biscuit-making wheats (Bowie and Rosella cultivars) differing in processing quality. Comparisons between these wheat cultivars were carried out on total wholemeal proteins, extracts with enriched starch granule proteins, and extracts enriched with gliadin storage proteins, with the intention of characterising, identifying, and cataloguing cultivar-specific proteins that could be used for segregation purposes. Initially, 2-dimensional gel electrophoresis was carried out on total wholemeal proteins using a broad range pH 3–10 immobilised pH gradient for the first dimension. Further screening was carried out using a combination of mid to narrow range immobilised pH gradients, including pH 4–7, 5.5–6.7, 5–8, 6–9, and 6–11. Best cultivar-specific protein fractionation was provided by the pH 5–8 range. Altogether, 4 unique cultivar-specific protein spots were excised from the pH 5–8 gels and identified by means of peptide mass fingerprinting, tandem mass spectrometry, or N-terminal sequencing. Starch granule protein extracts were prepared and fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A western blot was performed and probed with an anti-puroindoline-a antibody. Further to this, extracts enriched in gliadin storage proteins were isolated using 70% ethanol and analysed by 2-dimensional gel electrophoresis. The resulting gliadin protein maps showed 18 unique cultivar-specific gliadins. They were excised from the pH 6–9 gels and submitted for N-terminal amino acid sequencing. Overall, this study identified 23 proteins that could be used to distinguish between these closely related cultivars and may provide information on the molecular basis for the differences in processing exhibited by these wheats. The findings reported also contribute to a longer term objective of developing a broad and comprehensive knowledge base of commercial wheats, in regard to protein composition and their inherent processing qualities.

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Application of chromosome-specific monoclonal antibodies in wheat genetics

Genome ◽

10.1139/g92-126 ◽

1992 ◽

Vol 35 (5) ◽

pp. 831-837 ◽

Cited By ~ 2

Author(s):

R. E. Knox ◽

N. K. Howes ◽

T. Aung

Keyword(s):

Monoclonal Antibodies ◽

Gel Electrophoresis ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Seed Storage ◽

Seed Storage Proteins ◽

Polyacrylamide Gel ◽

Root Tip ◽

Chromosome Marker

A monoclonal antibody (P24B) to a wheat gliadin protein coded by a gene on the short arm of chromosome 1B was used as a chromosome marker. Somatic chromosome number for the 1B chromosome, as predicted by the level of binding of the antibody to extracts from seeds of single cross F2 and testcross F1 populations, was confirmed with sodium dodecyl sulfate – polyacrylamide gel electrophoresis and root-tip analysis. The implications of monoclonal antibodies as tools for cytogenetic analysis are discussed.Key words: polyacrylamide gel electrophoresis, aneuploid, cytogenetics, seed storage proteins.

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Precursors of storage proteins in Lupinus angustifolius

Biochemical Journal ◽

10.1042/bj2210333 ◽

1984 ◽

Vol 221 (2) ◽

pp. 333-341 ◽

Cited By ~ 8

Author(s):

K R Gayler ◽

B G Boadle ◽

M Snook ◽

E D Johnson

Keyword(s):

Protein Synthesis ◽

Western Blot ◽

Blot Analysis ◽

Gel Electrophoresis ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Lupinus Angustifolius ◽

Total Soluble Protein

The proteins that are synthesized during differentiation and development in the cotyledons of Lupinus angustifolius L. were characterized both in situ and after purification. The proteins present in situ were separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and subjected to ‘Western’-blot analysis to identify immunologically related polypeptides. The major storage proteins of the lupin, conglutins alpha and beta, were both present in juvenile tissue only as higher Mr precursors. For conglutin beta, a family of at least three polypeptides of Mr 66 000-72 000 accumulated during the earliest phases of protein synthesis in the developing cotyledon (20-28 days after flowering). Later in development each of these polypeptides disappeared and there was the concurrent appearance in the cotyledon of the lower-Mr fragments characteristic of mature conglutin beta. For conglutin alpha, an equivalent family of precursor polypeptides of Mr 60 000-83 000 was detected. Multiple internal sites for proteolytic cleavage of all these precursors appeared to be present. However, processing of the precursors was sufficiently slow to allow them to accumulate to over 50% of total soluble protein in juvenile tissue. The precursors were purified by column chromatography under non-dissociating conditions and shown by ultracentrifugation to be multimeric proteins with Mr values in the range 150 000-200 000.

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Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646313 ◽

1992 ◽

Vol 68 (05) ◽

pp. 534-538 ◽

Cited By ~ 5

Author(s):

Nobuhiko Yoshida ◽

Shingi Imaoka ◽

Hajime Hirata ◽

Michio Matsuda ◽

Shinji Asakura

Keyword(s):

Molecular Weight ◽

Sodium Dodecyl Sulfate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Tetraacetic Acid ◽

Fibrin Monomer ◽

Sds Page ◽

Thrombotic Tendency ◽

Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

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Low Molecular Weight Trypsin-Plasmin Inhibitors Isolated from Papain Treated Urinary Trypsin Inhibitor

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657115 ◽

1982 ◽

Vol 47 (01) ◽

pp. 014-018 ◽

Cited By ~ 13

Author(s):

H Sumi ◽

N Toki ◽

S Takasugi ◽

S Maehara ◽

M Maruyama ◽

...

Keyword(s):

Trypsin Inhibitor ◽

Gel Electrophoresis ◽

Specific Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Chromatography ◽

Urinary Trypsin Inhibitor ◽

Plasmin Activity ◽

Molecular Weights ◽

Plasmin Inhibitors

SummaryPapain treatment of human urinary trypsin inhibitor (UTI67; mol. wt. 43,000 by SDS-polyacrylamide gel electrophoresis, specific activity 1,897 U/mg protein) produced four new protease inhibitors, which were highly purified by gel chromatography on Sephadex G-100 and isoelectric focusing. The purified inhibitors (UTI26, UTI9-I, UTI9-II, and UTI9-III) were shown to be homogeneous by polyacrylamide disc gel electrophoresis, and had apparent molecular weights of 26,000, 9,000, 9,000, and 9,800, respectively, by sodium dodecyl sulfate gel electrophoresis. During enzymatic degradation of UTI67, the amino acid compositions changed to more basic, and the isoelectric point increased from pH 2.0 (UTI67) to pHs 4.4, 5.2, 6.6, and 8.3 (UTI26, UTI9-I, UTI9-II, and UTI9-III), respectively. Both the parent and degraded inhibitors had anti-plasmin activity as well as antitrypsin and anti-chymotrypsin activities. Much higher anti-plasmin/anti-trypsin and anti-plasmin/anti-chymotrypsin activities were observed in the degraded inhibitors than in the parent UTI67. They competitively inhibited human plasmin with Ki values of 1.13 X 10-7 - 2.12 X 10-6 M (H-D-Val-Leu-Lys-pNA substrate). The reactions were very fast and the active site of the inhibitors to plasmin was thought to be different from that to trypsin or chymotrypsin.

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