Significant immunological cross‐reactivity of plant glycoproteins

Laine, Anne‐Catherine; Faye, Loı̈c

doi:10.1002/elps.1150091210

Cited by 28 publications

(12 citation statements)

References 7 publications

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“…B, Purified native AtPAP26 from 2Pi Arabidopsis suspension cells (50 ng/lane; Veljanovski et al, 2006) and clarified extract proteins from shoots (2 mg/lane) and roots (4 mg/lane) of the +Pi and 2Pi seedlings were resolved by SDS-PAGE and electroblotted onto a poly(vinylidene difluoride) membrane. Following oxidation of antigenic glycosyl groups with sodium-mperiodate (Laine, 1988), blots were probed with a 1,000-fold dilution of anti-(native AtPAP26)-immune serum and immunoreactive polypeptides detected using an alkaline-phosphatase-linked secondary antibody and chromogenic detection (Veljanovski et al, 2006). C, APase activity of clarified extracts represent means (6SE) of duplicate assays on n = 3 biological replicates; asterisks denote values that are significantly different from Col-0 (P , 0.01).…”

Section: Atpap26 Is the Predominant Intracellular And A Major Secretementioning

confidence: 99%

The Dual-Targeted Purple Acid Phosphatase Isozyme AtPAP26 Is Essential for Efficient Acclimation of Arabidopsis to Nutritional Phosphate Deprivation

et al. 2010

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Induction of intracellular and secreted acid phosphatases (APases) is a widespread response of orthophosphate (Pi)-starved (2Pi) plants. APases catalyze Pi hydrolysis from a broad range of phosphomonoesters at an acidic pH. The largest class of nonspecific plant APases is comprised of the purple APases (PAPs). Although the biochemical properties, subcellular location, and expression of several plant PAPs have been described, their physiological functions have not been fully resolved. Recent biochemical studies indicated that AtPAP26, one of 29 PAPs encoded by the Arabidopsis (Arabidopsis thaliana) genome, is the predominant intracellular APase, as well as a major secreted APase isozyme up-regulated by 2Pi Arabidopsis. An atpap26 T-DNA insertion mutant lacking AtPAP26 transcripts and 55-kD immunoreactive AtPAP26 polypeptides exhibited: (1) 9-and 5-fold lower shoot and root APase activity, respectively, which did not change in response to Pi starvation, (2) a 40% decrease in secreted APase activity during Pi deprivation, (3) 35% and 50% reductions in free and total Pi concentration, respectively, as well as 5-fold higher anthocyanin levels in shoots of soil-grown 2Pi plants, and (4) impaired shoot and root development when subjected to Pi deficiency. By contrast, no deleterious influence of AtPAP26 loss of function occurred under Pi-replete conditions, or during nitrogen or potassium-limited growth, or oxidative stress. Transient expression of AtPAP26-mCherry in Arabidopsis suspension cells verified that AtPAP26 is targeted to the cell vacuole. Our results confirm that AtPAP26 is a principal contributor to Pi stress-inducible APase activity, and that it plays an important role in the Pi metabolism of 2Pi Arabidopsis.Orthophosphate (Pi) is an essential plant macronutrient required for many pivotal metabolic processes such as photosynthesis and respiration. However, the massive use of Pi fertilizers in agriculture demonstrates how the free Pi level of many soils is suboptimal for plant growth. The world's reserves of rock phosphate, our major source of Pi fertilizers, are projected to be depleted by the end of this century (Vance et al., 2003). Furthermore, Pi runoff from fertilized fields into nearby surface waters results in environmentally destructive processes such as aquatic eutrophication and blooms of toxic cyanobacteria. Effective biotechnological strategies are needed to engineer Pi-efficient transgenic crops to ensure agricultural sustainability and a reduction in Pi fertilizer overuse. This necessitates a detailed understanding of Pi-starvation-inducible (PSI) gene expression and the complex morphological, physiological, and biochemical adaptations of Pi-deficient (2Pi) plants.A well-documented component of the plant Pi stress response is the up-regulation of intracellular and secreted acid phosphatases (APases; E.C. 3.1.3.2) that catalyze the hydrolysis of Pi from various phosphate monoesters and anhydrides in the acidic pH range (Tran et al., 2010a). APase induction by 2Pi plants has been correlated wit...

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Section: Atpap26 Is the Predominant Intracellular And A Major Secretementioning

confidence: 99%

The Dual-Targeted Purple Acid Phosphatase Isozyme AtPAP26 Is Essential for Efficient Acclimation of Arabidopsis to Nutritional Phosphate Deprivation

et al. 2010

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“…Some of the sidechains have been characterized and reach a level of complexity not previously described in other plant glycoproteins (see Faye eta/., 1989 for review;Takahashi eta/., 1986). It has been shown that plant complex glycans are highly immunogenic in rabbits (Laine and Faye, 1988). Therefore, purified sycamore cell laccase was submitted to chemical deglycosylation with TFMS prior to immunization.…”

Section: Discussionmentioning

confidence: 99%

Characterization and localization of laccase forms in stem and cell cultures of sycamore

Driouich

Laine

Vian³

et al. 1992

The Plant Journal

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SummaryA laccase-type polyphenoloxidase (EC 1.1 0.3.2.), abundantly secreted by suspension-cultured sycamore (Acer pseudoplatanus) cells was purified t o homogeneity. This laccase form is a glycoprotein (molecular weight 110 000) with high mannose and complex glycans. The polypeptide moiety has a molecular weight of 66 000, indicating that the glycoprotein is 40% carbohydrate. Laccase is abundantly present in both the cell wall and the culture medium of suspensioncultured sycamore cells, but it is not detected in the cytoplasm, indicating that this large protein is efficiently secreted by the cells. Polyclonal rabbit antiserum was raised against the deglycosylated protein and was used to probe extracts of sycamore stem tissues. A second laccase form (molecular weight 56 OOO), antigenically related t o laccase from cell cultures, is abundant in the epidermis of sycamore stems. In addition, this 56 kDa laccase form co-localizes with lignin precursors on tissue prints from sycamore stems. A polypeptide (molecular weight 50 000-56 000), antigenically related t o sycamore laccase, was also immunodetected in most plant organs previously described in the literature as polyphenoloxidase-rich.

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“…Controls for affinodetection were performed in the presence of 0.3 M e-methyl-mannoside (ConA). Controls for immunodetection were done using periodate (NaIO4) oxidation as described (Laine and Faye 1988).…”

Section: Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis (Smentioning

confidence: 99%

The role of high-mannose and complex asparagine-linked glycans in the secretion and stability of glycoproteins

Driouich

Gonnet

Makkie

et al. 1989

Planta

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Suspension-cultured cells of sycamore (Acer pseudoplatanus L.) secrete a number of acid hydrolases and other proteins that have both highmannose and complex asparagine-linked glycans. We used affinity chromatography with concanavalin A and an antiserum specific for complex glycans in conjunction with in vivo-labeling studies to show that all of the secreted proteins carry glycans. The presence of complex glycans on secretory proteins indicates that they are passing through the Golgi complex on the way to the extracellular compartment. The sodium ionophore, monensin, did not block the transport of proteins to the extracellular medium, even though monensin efficiently inhibited the Golgi-mediated processing of complex glycans. The inhibition of N-glycosylation by tunicamycin reduced by 76% to 84% the accumulation of newly synthesized (i.e. radioactively labeled) protein that was secreted by the sycamore cells, while cytoplasmic protein biosynthesis was not affected by this antibiotic. However, in the presence of glycoprotein-processing inhibitors, such as castanospermine and deoxymannojirimycin, the formation of complex glycans was prevented but glycoprotein secretion was unchanged. These results support the conclusion that N-linked glycan processing is not necessary for sorting, but glycosylation is required for accumulation of secreted proteins in the extracellular compartment.

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Significant immunological cross‐reactivity of plant glycoproteins

Cited by 28 publications

References 7 publications

The Dual-Targeted Purple Acid Phosphatase Isozyme AtPAP26 Is Essential for Efficient Acclimation of Arabidopsis to Nutritional Phosphate Deprivation

The Dual-Targeted Purple Acid Phosphatase Isozyme AtPAP26 Is Essential for Efficient Acclimation of Arabidopsis to Nutritional Phosphate Deprivation

Characterization and localization of laccase forms in stem and cell cultures of sycamore

The role of high-mannose and complex asparagine-linked glycans in the secretion and stability of glycoproteins

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