Secondary structure of oleosins in oil bodies isolated from seeds of safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.)

Oil bodies were isolated from mature seeds of sunflower (Helianthus annuus L.) and safflower (Carthamus tinctorius L.). Oil body preparations containing only oleosin proteins could be obtained from safflower seeds by salt-washing followed by centrifugation on discontinuous sucrose density gradients. However, it was necessary to treat sunflower oil bodies with urea to obtain preparations of similar purity. Incubation of the oil bodies with proteinases gave two fragments with molecular masses of 6 and 8 kDa which were protected from digestion. These fragments represented the hydrophobic domain of the oleosins, as determined by N-terminal sequencing. Intact and proteinase-treated oil bodies of both species were analysed by Fourier-transform infrared spectroscopy, as dry films and in aqueous medium, the spectra being compared with those obtained for pure oil samples in order to identify the bands resulting from the oleosin proteins and protected peptides. This investigation showed that the hydrophobic domain of the oleosins in intact oil bodies is predominantly α-helical in structure and that the conformation was not greatly affected by washing the oil bodies with urea during preparation.

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Purification and characterization of oil-bodies (oleosomes) and oil-body boundary proteins (oleosins) from the developing cotyledons of sunflower (Helianthus annuus L.)

Biochemical Journal ◽

10.1042/bj3140333 ◽

1996 ◽

Vol 314 (1) ◽

pp. 333-337 ◽

Cited By ~ 66

Author(s):

Mark MILLICHIP ◽

Arthur S. TATHAM ◽

Frances JACKSON ◽

Gareth GRIFFITHS ◽

Peter R. SHEWRY ◽

...

Keyword(s):

Helianthus Annuus ◽

Storage Proteins ◽

Phospholipid Content ◽

Major Protein ◽

Oil Body ◽

Oil Bodies ◽

Helianthus Annuus L ◽

Novel Structure ◽

Body Boundary ◽

Helical Proteins

Oil-bodies, from the immature cotyledons of sunflower (Helianthus annuus L.), were difficult to purify to homogeneity using conventional techniques. The major protein contaminants were albumin and globulin storage proteins. A protocol has been developed, therefore, based upon the stringent washing of the oil-body fraction in 9 M urea, which effectively removed almost all the contaminating protein as judged by SDS/PAGE. The urea-washed oil-bodies were enriched in two major proteins of Mr 19000 and 20000. These proteins were oleosins as demonstrated by their amino acid compositions and the sequence analysis of peptides produced by CNBr cleavage. Far-UV CD spectra of the oleosins in trifluoroethanol, trifluoroethanol/water mixtures and as mixed micelles in SDS, were typical of α-helical proteins with α-helical contents of some 55%. The phospholipid content of the urea-washed preparations was less than 0.1% of that required to form a half-unit membrane surrounding the oil-body. The oil-body surface therefore appears to be an unusual and novel structure, covered largely by an oleosin protein coat or pellicle rather than a conventional fluid membrane, half-unit or otherwise.

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The utilisation of fatty-acid substrates in triacylglycerol biosynthesis by tissue-slices of developing safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.) cotyledons

Planta ◽

10.1007/bf00401017 ◽

1988 ◽

Vol 173 (3) ◽

pp. 309-316 ◽

Cited By ~ 17

Author(s):

Gareth Griffiths ◽

Sten Stymne ◽

A. Keith Stobart

Keyword(s):

Fatty Acid ◽

Helianthus Annuus ◽

Carthamus Tinctorius ◽

Tissue Slices ◽

Helianthus Annuus L ◽

Triacylglycerol Biosynthesis

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Oil-bodies from sunflower (Helianthus annuus L.) seeds

Biochemical Journal ◽

10.1042/bj3170955 ◽

1996 ◽

Vol 317 (3) ◽

pp. 955-956 ◽

Cited By ~ 8

Author(s):

Frédéric BEISSON ◽

Nathalie FERTE ◽

Georges NOAT

Keyword(s):

Helianthus Annuus ◽

Oil Bodies ◽

Helianthus Annuus L

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SALINITY TOLERANCE CLASSIFICATION OF SUNFLOWER (Helianthus annuus L.) AND SAFFLOWER (Carthamus tinctorius L.) BY CLUSTER AND PRINCIPAL COMPONENT ANALYSIS

Applied Ecology and Environmental Research ◽

10.15666/aeer/1702_38493857 ◽

2019 ◽

Vol 17 (2) ◽

pp. 3849-3857 ◽

Cited By ~ 3

Author(s):

M D KAYA ◽

G AKDOĞAN ◽

E G KULAN ◽

H DAĞHAN ◽

A SARI

Keyword(s):

Principal Component Analysis ◽

Helianthus Annuus ◽

Salinity Tolerance ◽

Principal Component ◽

Component Analysis ◽

Carthamus Tinctorius ◽

Helianthus Annuus L

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The activity of antioxidant enzymes in response to salt stress in safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.) seedlings raised from seed treated with chitosan

Journal of the Science of Food and Agriculture ◽

10.1002/jsfa.5953 ◽

2012 ◽

Vol 93 (7) ◽

pp. 1699-1705 ◽

Cited By ~ 34

Author(s):

Nusrat Jabeen ◽

Rafiq Ahmad

Keyword(s):

Antioxidant Enzymes ◽

Salt Stress ◽

Helianthus Annuus ◽

Carthamus Tinctorius ◽

Helianthus Annuus L

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Some studies on the composition and surface properties of oil bodies from the seed cotyledons of safflower (Carthamus tinctorius) and linseed (Linum ustatissimum)

Biochemical Journal ◽

10.1042/bj1900551 ◽

1980 ◽

Vol 190 (3) ◽

pp. 551-561 ◽

Cited By ~ 68

Author(s):

C R Slack ◽

W S Bertaud ◽

B D Shaw ◽

R Holland ◽

J Browse ◽

...

Keyword(s):

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Electron Microscopic Examination ◽

Carthamus Tinctorius ◽

Oil Body ◽

Oil Bodies ◽

Electron Microscopic ◽

Intact Cells ◽

Glycerolipid Composition ◽

Induced Aggregation

1. The average oil-body diameter in intact cells of developing linseed (Linum usitatissimum) and safflower (Carthamus tinctorius) cotyledons was similar (about 1.4 micrometer), and there was little change in size after oil bodies were isolated and repeatedly washed. 2. The glycerolipid composition of washed oil bodies from both developing and mature cotyledons of the two species was similar; oil bodies from ten different batches of cotyledons contained 4.3 +/- 0.16 mumol of 3-sn-phosphatidylcholine and 25.2 +/- 1.7 mumol of diacylglycerol per 1000 mumol of triacylglycerol. During four successive washings of a once-washed oil-body preparation, the proportion of diacylglycerol to triacylglycerol remained constant and that of 3-sn-phosphatidylcholine to triacylglycerol decreased by only 20%. 3. The protein content of thrice-washed oil bodies from the two species was similar, about 2.4% of the weight of glycerolipids, and appeared to be independent of the stage of cotyledon maturity. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated that the protein of purified oil bodies from the two species consisted mainly of only four polypeptides and that two of the polypeptides from each species had apparent mol.wts. of 17500 and 15500. Similar patterns of polypeptides were obtained after the hydrolysis of the 15500-mol.wt. polypeptides from linseed and safflower oil bodies by Staphylococcus aureus V8 proteinase, whereas the proteolysis of the 17500-mol.wt. polypeptides from the two species produced different patterns of polypeptides. 4. The 3-sn-phosphatidylcholine in oil-body preparations was hydrolysed about 85% by bee-venom phospholipase A2 without any apparent coalescence of the oil bodies. Incubation with lipase from Rhizopus arrhizus caused rapid coalescence of the oil bodies, and this lipase appeared to initially hydrolyse diacylglycerols in preference to triacylglycerol. 5. Oil bodies from both species were almost completely dispersed in suspensions of pH between 7.1 and 8.3, but formed large aggregates at pH values between 6.7 and 3.9; pH-induced aggregation caused no coalescence. Aggregates formed under acidic conditions were dispersed by re-adjusting the pH of suspensions to 8.3. 6. A freeze-etch electron-microscopic examination of isolated oil bodies indicated that these organelles were bounded by some form of membrane with a particle-free outer surface.

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