Immunochemical crossreactivity between zein, hordein and gliadin

Dierks-Ventling, Christa; Cozens, Karen

doi:10.1016/0014-5793(82)80239-1

Cited by 31 publications

(10 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some antibodies reacted with one or two polypeptides only (CMpXhor, CMpVhor, CMpYhor), where- The recognition of a number of polypeptides by a given MAb gives further evidence for primary structure homology among hordein polypeptide. Employing polyclonal antibodies to hordein polypeptides these homologies have been implicated and homologies among prolamins from different cereal species have been demonstrated (10,16,17,45). The broad specificity within (CMpBhorI) and among (CMpBChor) hordein groups is probably due to the location of the epitopes within repetitive glutamine + proline blocks which are common to the B-and C-hordein polypeptides (19,37).…”

Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies to hordein polypeptides

Ullrich

Rasmussen

Høyer‐Hansen

et al. 1986

Carlsberg Res. Commun.

View full text Add to dashboard Cite

Monoclonal antibodies to hordein polypeptides have been produced and characterized. All antibodies reacted with more than one polypeptide, indicating that different hordein polypeptides have common epitopes. Of the seventeen isolated hybridoma lines, six secreted antibodies which reacted both with B-and C-hordein, five recognized only B-hordein and one secreted antibodies specific for C-hordein polypeptides. Two clones produced antibodies to an unidentified polypeptide doublet with an apparent molecular weight of 43 kD. These two polypeptides have at least one epitope in common with B-hordein polypeptides and are not extractable in water but in alcohol. They are not visible by Coomassie blue staining.The mutant M56 fails to synthesize B-hordein polypeptides due to a major deletion. It contains polypeptides in the B-hordein region of the gel, which have epitopes recognized by the B-hordein antibodies, but are different from B-hordeins. The 43 kD hordein doublet was more abundant in the mutant than in the wild-type endosperm, suggesting that these polypeptides like the C-hordein polypeptides are synthesized in larger amounts as a consequence of the absent B-hordein polypeptide synthesis. Mutant 1508 which has an impaired synthesis of both B-and C-hordein polypeptides was also found to be defective in the synthesis of the 43 kD polypeptide doublet. It was shown that immunofluoreseent techniques may be of practical importance to identify mutants with elevated or reduced ~tmounts of specific hordein polypeptides. Due to the higher sensitivity compared to SDS-PAGE, immunoblotting can be a useful tool for genotype identification in breeding programmes.

show abstract

Section: Discussionmentioning

confidence: 99%

Monoclonal antibodies to hordein polypeptides

Ullrich

Rasmussen

Høyer‐Hansen

et al. 1986

Carlsberg Res. Commun.

View full text Add to dashboard Cite

show abstract

“…7). It has also been shown that purified anti-zein IgG cross-reacted with hordeins, but that anti-hordein IgG did not cross-react with zeins, and that both cross-reacted with a few proteins of the gliadin family [46].…”

Section: Discussionmentioning

confidence: 99%

Antibodies to the photosystem I chlorophyll a+b antenna cross-react with polypeptides of CP29 and LHCII

White

Green

1987

Eur J Biochem

View full text Add to dashboard Cite

A chlorophyll (a + b)-protein complex associated with photosystem I (PSI) was isolated from a larger PSI complex (CPIa) produced by electrophoresis of barley thylakoids solubilized with 300 mM octyl glucoside. It had an apparent M , of 35000-43000 on 7.5% and 10% acrylamide gels respectively, and a chlorophyll a/b ratio of 2.5 f 1.5. Denaturation released four polypeptides migrating between 21 -24 kDa. They were well separated from the polypeptides of the two photosystem I1 chlorophyll a + b antenna complexes: LHCII (25 -27 kDa) and CP29 (28 -29 kDa).In order to study the PSI antenna complex, antibodies were raised against highly purified CPIa. The antigen appeared to be pure when electrophoresed, blotted and reacted with its antiserum, i.e. anti-CPIa detected only the 64-66-kDa CPI apoprotein and the four 21 -24 kDa antenna polypeptides. However, when blotted against the whole spectrum of thylakoid proteins, it cross-reacted with both LHCII and CP29 apoproteins. Removal of anti-CPI activity from the anti-CPIa did not affect these cross-reactions, showing that they were not due to antibodies directed against CPI. To show that the same antibody population was reacting with both the photosystem I and photosystem I1 antenna polypeptides, anti-CPIa was adsorbed onto highly purified CPIa on nitrocellulose. The bound antibody was eluted and used again in a Western blot against whole thylakoid proteins. This selected antibody population showed the same relative strength of reaction with photosystem I and photosystem I1 antenna polypeptides as the original antibody population had.Similar observations have been made with antibodies to the two photosystem I1 antenna complexes. We therefore conclude that there are antigenic determinants in common among the chlorophyll a + b binding polypeptides, and predict that there could be amino acid sequence similarities.All the chlorophyll (Chl) in thylakoid membranes is bound to specific proteins [I] to form chlorophyll-protein (CP) complexes. There are three Chl-a-containing complexes known as CPI, CP47 and CP43. They are closely associated with reaction centers and their polypeptides are synthesized in the chloroplast [2-61. There are also at least three Chla+b-containing complexes known as LHCII, CP29 and the PSI antenna complex or LHCI (see [7 -91) antenna (Chl a/b = 3-4) associated with PSI [S, 161. There are several versions of LHCI (or PSI antenna) containing between one and four polypeptides, depending on the isolation method used [9,[17][18][19][20][21][22][23][24]. In this paper, a simple method requiring only octyl glucoside solubilization and gel electrophoresis is used to isolate a PSI antenna-CP complex containing four polypeptides in the 21 -24-kDa range. It is isolated from CPIa, a PSI complex which consists of CPI (the P700 reaction center complex of PSI) and the PSI antenna complex. Polyclonal antibodies to this complex were found to cross-react with polypeptides of the two PSII Chl a + b protein complexes, LHCII and CP29. MATERIALS AND METHODSPreparation and deter...

show abstract

“…Identification of Zein by Immumoblotting. Zein was identified by an immunoblotting method as described previously (4). In brief, proteins separated by gel electrophoresis were immediately transferred electrophoretically to nitrocellulose sheets (28 Plant Physiol.…”

mentioning

confidence: 99%

Expression of Zein in Long Term Endosperm Cultures of Maize

Shimamoto¹,

Ackermann²,

Dierks-Ventling³

1983

Plant Physiol.

Self Cite

View full text Add to dashboard Cite

Continuous cultures, established 10 days after pollination from endosperms of inbred A636 Zea mays (L.) were extracted 21 months later with aqueous ethanol. The solubilized proteins were analyzed by polyacrylamide-sodium dodecyl sulfate gel electrophoresis. Two protein bands co-migrated with zein, the major storage protein of maize. Immunoblotting of the gel followed by incubation of the immobilized proteins with anti-zein IgG provided evidence that the polypeptides were in fact zein. Electron microscopic studies showed that the cultures contained cells with protein bodies as found in developing endosperms. The protein bodies could be isolated from the cultures and were shown to contain zein. We conclude that the long term cultures described here synthesize zein and deposit it in the form of protein bodies of the type found in developing endosperms. Thus, certain endosperm characteristics and the production of tissue-specific proteins are retained in prolonged culture.Tissue cultures which express tissue-specific proteins are highly desirable for studying the regulatory mechanisms leading to the differential expression of these proteins. In higher plants such culture systems have, to our knowledge, not been described, mainly because few plant proteins have been studied for their tissue specificity. Among the better known ones are storage proteins of seeds accumulating in developing endosperms of cereal grains (14) or in cotyledons of legumes (17). Attempts to establish long term cultures of cereal endosperms have been described (9). The first report of a successful, continuous culture of maize endosperm appeared in 1949 (16). Several more reports on endosperm cultures followed some ofwhich have been shown to produce anthocyanin (7, 25) and starch (2). Production of storage proteins in such cultures has not been investigated. Zein, which constitutes more than 50% of the seed storage proteins in maize, is synthesized on membrane-bound polysomes and deposited in the form ofprotein bodies (1, 1 1). Immunohistochemical studies have shown that zein is specifically localized in the endosperm (5). Because of its tissue specifity and the rapidly expanding knowledge of its biochemistry and molecular biology (8,13,14,20) Initiation and Maintenance of Endosperm Cultures. After carefully removing all husks and silks, ears were surface-sterilized in 0.01% mercuric chloride for 20 min and rinsed three times with sterile distilled H20. The tops of the 10-d-old kernels were cut off with a scalpel, and the endosperms were scooped out with forceps carefully avoiding the embryo. Isolated endosperms were immediately placed on 6-cm-diameter plastic Petri dishes containing 10 ml agar medium and the dishes were sealed with Parafilm. Strauss medium (26) modified according to Shannon and Batey (23) to contain 2 g/l L-asparagine and 2 g/l yeast extracts (Merck, Munich, F.R.G.) was used. All the cultures were grown in the dark at 25°C and subcultured every 3 to 4 weeks.Electron Microscopy. Two mm3 sections of each culture were pr...

show abstract

Immunochemical crossreactivity between zein, hordein and gliadin

Cited by 31 publications

References 15 publications

Monoclonal antibodies to hordein polypeptides

Monoclonal antibodies to hordein polypeptides

Antibodies to the photosystem I chlorophyll a+b antenna cross-react with polypeptides of CP29 and LHCII

Expression of Zein in Long Term Endosperm Cultures of Maize

Contact Info

Product

Resources

About