Structure of maize protein bodies and immunocytochemical localization of zeins

Lending, Craig R.; Kriz, Alan L.; Larkins, Brian A.; Bracker, Charles E.

doi:10.1007/bf01282959

Cited by 111 publications

(66 citation statements)

References 24 publications

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“…7, A and C). The protein bodies were lined by a dark staining matrix and had a central light staining chamber very similar to the protein bodies in corn endosperm (Lending et al, 1988). In some sections, the whole cluster of protein bodies appeared to be lined by the RER (Fig.…”

Section: Localization Of the 15-kd Zein Protein In Leaf Cells Of Tranmentioning

confidence: 76%

“…The nitrocellulose was blocked overnight with 1% BSA in Tris-buffered saline containing 0.0576 Tween 20 and was then incubated in the same solution with the appropriate antibody. The anti-15-kD zein an ibody was kindly provided by Dr. Brian Larkins (Lending et al, 1988). The protein bands reacting to the antibodies were made visible using an alkaline phosphatase-linked second antibody and the substrates nitroblue tetrazolium and 5-bromo-4-chloro-3-indolyl-phosphate according to the manufacturer's instructions (Promega).…”

Section: Western Analysismentioning

confidence: 99%

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Accumulation of 15-Kilodalton Zein in Novel Protein Bodies in Transgenic Tobacco

et al. 1995

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Zeins, the seed storage proteins of maize, are a group of alcoholsoluble polypeptides of different molecular masses that share a similar amino acid composition but vary i n their sulfur amino acid composition. They are synthesized on the rough endoplasmic reticulum (ER) in the endosperm and are stored in ER-derived protein bodies. Our goal i s to balance the amino acid composition of the methionine-deficient forage legumes by expressing the sulfur amino acid-rich 15-kD zeins in their leaves. However, it is crucial to know whelher this protein would be stable i n nonseed tissues of transgenic plants. The major focus of this paper is to compare the accumulation pattern of the 15-kD zein protein with a vacuolar targeted seed protein, p-phaseolin, in nonseed tissues and to determine the basis for i t s stability/instability. We have introduced the 15-kD zein and bean p-phaseolin-coding sequences behind the 35s cauliflower mosaic virus promoter into tobacco (Nicofiana fabacum) and analyzed the protein's accumulation pattern in different tissues. Our results demonstrate that the 15-kD seed protein i s stable not only i n seeds but in all nonseed tissues tested, whereas the P-phaseolin protein accumulated only i n mid-and postmaturation seeds. Interestingly, zein accumulates in novel protein bodies both in the seeds and in nonseed tissues. We attribute the instability of the p-phaseolin protein i n nonseed tissues to the fact that it is targeted to protease-rich vacuoles. The stability of the 15-kD zein could be attributed to its retention i n the ER or to the proteaseresistant nature of the protein.Seed storage proteins constitute a potentially useful class of proteins for the improvement of forage crops if they can be made to accumulate in leaves. It is a fairly simple genetic engineering feat to introduce a seed protein-coding sequence behind a strong constitutive promoter into transgenic plants and ensure high rates of synthesis of the corresponding protein. However, stability of the protein in an alien environment is still not clearly defined and has to be treated on a case by case basis.Seed proteins are synthesized during seed development and accumulate in protein bodies. These proteins are then utilized by the emerging seedling during germination. The major seed protein in dicotyledonous plants are the saltsoluble globulins, which are stored in vacuole-derived protein bodies. Most monocot seed storage proteins are '

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Section: Localization Of the 15-kd Zein Protein In Leaf Cells Of Tranmentioning

confidence: 76%

Section: Western Analysismentioning

confidence: 99%

Accumulation of 15-Kilodalton Zein in Novel Protein Bodies in Transgenic Tobacco

et al. 1995

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“…For immunoblot analysis, proteins were electroblotted from 10% SDS-polyacrylamide gels to nitrocellulose using a semidry blotting apparatus (Trans-Blot SD, Bio-Rad) and the buffer system described by Bjerrum and Schafer-Nielsen (1986). Filters were probed with polyclonal antiserum (1:5000 dilution) directed against a-zeins and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5000 dilution; Lending et al, 1988). Immunoblots were developed with detection reagents provided with a western blotting kit (ECL, Amersham) according to the manufacturer's instructions.…”

Section: Fluorography and Immunoblot Analysismentioning

confidence: 99%

“…The processed f/Z a-zein (ValbAla) was localized in the soluble portion of the microsomes, whereas the f/Z a-zein co-fractionated with the microsomal membranes. By remaining anchored t o protein body membranes during endosperm maturation, the f/Z zein may thus constrain storage protein packing and perturb protein body morphology.The maize (Zea mays L.) floury-2 fl2) mutation produces pleiotropic effects that include a soft, starchy endosperm, a reduction in the amount of endosperm storage protein, and changes in the arrangement of proteins within protein bodies (Nelson et al, 1965;Lending et al, 1988). The changes in protein packaging are accompanied by an accumulation of aberrant protein bodies near the nuclei of endosperm cells.…”

mentioning

confidence: 99%

“…The maize (Zea mays L.) floury-2 fl2) mutation produces pleiotropic effects that include a soft, starchy endosperm, a reduction in the amount of endosperm storage protein, and changes in the arrangement of proteins within protein bodies (Nelson et al, 1965;Lending et al, 1988). The changes in protein packaging are accompanied by an accumulation of aberrant protein bodies near the nuclei of endosperm cells.…”

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A Defective Signal Peptide Tethers the floury-2 Zein to the Endoplasmic Reticulum Membrane

et al. 1997

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The maize (Zea mays L.) floury-2 (f/Z) mutation is associated with a general decrease in storage protein synthesis, altered protein body morphology, and the synthesis of a novel 24-kD a-zein storage protein. Unlike storage proteins in normal kernels and the majority of storage proteins in f/Z kernels, the 24-kD a-zein contains a signal peptide that would normally be removed during protein synthesis and processing. The expected processing site of this a-zein reveals a putative mutation alanine-valine (Ala-Val) that is not found at other junctions between signal sequences and mature proteins. To investigate the impact of such a mutation on signal peptide cleavage, we have assayed the 24-kD f/Z a-zein in a co-translational processing system i n vitro. Translation of RNA from f/Z kernels or synthetic RNA encoding the f/Z a-zein in the presence of microsomes yielded a 24-kD polypeptide. A normal signal peptide sequence, generated by site-directed mutagenesis, restored the capacity of the RNA to direct synthesis of a properly processed protein i n a cell-free system. Both the f/Z a-zein and the f/Z a-zein (Val-Ala) were translocated into the lumen of the endoplasmic reticulum. The processed f/Z a-zein (ValbAla) was localized in the soluble portion of the microsomes, whereas the f/Z a-zein co-fractionated with the microsomal membranes. By remaining anchored t o protein body membranes during endosperm maturation, the f/Z zein may thus constrain storage protein packing and perturb protein body morphology.The maize (Zea mays L.) floury-2 fl2) mutation produces pleiotropic effects that include a soft, starchy endosperm, a reduction in the amount of endosperm storage protein, and changes in the arrangement of proteins within protein bodies (Nelson et al., 1965;Lending et al., 1988). The changes in protein packaging are accompanied by an accumulation of aberrant protein bodies near the nuclei of endosperm cells. Coincident with the appearance of the aberrant protein bodies is an increase in the accumulation

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Sorting of Storage Proteins in the PlantGolgi Apparatus

Hinz

Herman²

2018

Annual Plant Reviews Online

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The sections in this article areIntroductionSeed Storage Proteins and Ancillary Proteins are Stored in Two Different Types of Storage Organelles, Protein Bodies and Protein Storage VacuolesOrigin ofPSVsand Relationship to Lytic VacuoleGolgi‐Mediated Sorting of Storage Proteins into thePSVsVacuolar Sorting Signals are Recognized by aGolgi‐Localized Sorting MachineryGolgi Apparatus Transport Vesicles in SeedsPutative Sorting MechanismsVacuolar Sorting Pathways are not Exclusive for a Certain Cell TypeAutophagy Constitutes an Alternative, Novel Route Bypassing Progression through theGolgi Apparatus for the Vacuolar Accumulation of Storage Proteins during Seed MaturationPerspectives of Future Research

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Structure of maize protein bodies and immunocytochemical localization of zeins

Cited by 111 publications

References 24 publications

Accumulation of 15-Kilodalton Zein in Novel Protein Bodies in Transgenic Tobacco

Accumulation of 15-Kilodalton Zein in Novel Protein Bodies in Transgenic Tobacco

A Defective Signal Peptide Tethers the floury-2 Zein to the Endoplasmic Reticulum Membrane

Sorting of Storage Proteins in the PlantGolgi Apparatus

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