Production and Characterization of Monoclonal Antibodies to Wall-Localized Peroxidases from Corn Seedlings

Kim, Sung-Ha; Terry, Maurice E.; Hoops, Pepper; Dauwalder, Marianne; Roux, Stanley J.

doi:10.1104/pp.88.4.1446

Cited by 29 publications

(17 citation statements)

References 22 publications

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“…were transplanted into a commercial potting soil (12 tillers per 25-cm-diameter pot) and grown in a controlled-environment chamber at a mean photon flux density of 500 ,gmol mr2 s-' PAR (24); details of extraction procedures are described in the following section. The duration of vacuum infiltration used to extract apoplastic fluid from tissue segments has commonly ranged from 30 s to 4 min (4,14). However, Li et al.…”

Section: Materials and Methods Plant Materials And Growth Measurementsmentioning

confidence: 99%

Peroxidase Activity in the Leaf Elongation Zone of Tall Fescue

MacAdam¹,

Sharp²,

Nelson³

1992

Plant Physiol.

110

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Previous work suggested that cell wall peroxidase activity increased as cells were displaced through the elongation zone in leaf blades of tall fescue (Festuca arundinacea Schreb.). In this study, two genotypes that differ in length of the elongation zone were used to examine the relationship between peroxidase activity in apoplastic fluid of intact leaf blade segments and the spatial distribution of leaf growth. Apoplastic fluid was extracted by vacuum infiltration and centrifugation, and peroxidase activity was assayed spectrophotometrically. Isoelectric focusing was used to characterize the isoforms of apoplastic peroxidase within the region of elongation and in the region of secondary cell wall deposition, which is distal to the elongation zone. A striking correlation was found in each genotype between both the location and timing of increase in apoplastic peroxidase activity and the onset of growth deceleration. Only cationic isoforms of apoplastic peroxidase could be identified in the elongation zone, whereas additional anionic isoforms appeared in the region of secondary cell wall deposition. We conclude that cessation of elongation growth in tall fescue leaf blades is likely to be related to the secretion of cationic isoforms of peroxidase into the cell wall.Cessation of elongation occurs rapidly as cells are displaced through the distal half of the elongation zone of tall fescue (Festuca arundinacea Schreb.) leaf blades (18). The wall tightening that is thought to decrease the rate of cell expansion may result from formation of cross-linkages among cell wall polymers. Many of these covalent bonds occur by oxidation/ reduction reactions catalyzed by peroxidase (7).In the companion paper (17), we examined the spatial and temporal relationships between ionically bound peroxidase activity, which has previously been equated with cell wall activity (8,10,21), and the profile of elongation growth in two genotypes of tall fescue differing in length of the elongation zone.

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Section: Materials and Methods Plant Materials And Growth Measurementsmentioning

confidence: 99%

Peroxidase Activity in the Leaf Elongation Zone of Tall Fescue

MacAdam¹,

Sharp²,

Nelson³

1992

Plant Physiol.

110

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Section: Methodsmentioning

confidence: 99%

“…3, the R and far-red light (FR) sources (source A) used were those described by Chen and Roux (11). Fluence rates of R were [8][9][10][11][12] jtmol/m2 sec; fluence rates for FR were 15-20 ,4mol/m2 sec.For the data in Fig. 3, R-irradiations were given in a 2-min pulse at 10-20 gmol/m2sec and measured by a quantum sensor (LI-185B; Li-cor, Lincoln, NE) by using a Schott KL1500 fiberoptic microscope illuminator (source B) filtered with a Schott RG610 filter.…”

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confidence: 99%

Phytochrome induces changes in the immunodetectable level of a wall peroxidase that precede growth changes in maize seedlings

Kim

Shinkle

Roux

1989

Proc. Natl. Acad. Sci. U.S.A.

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The regulatory pigment phytochrome induces rapid and opposite growth changes in different regions of etiolated maize seedlings: it stimulates the elongation rate of coleoptiles and inhibits that of mesocotyls. As measured by a quantitative immunoassay, phytochrome also promotes rapid and opposite changes in the extractable content of a Mr 98,000 anionic isoperoxidase in the cell walls of these same organs: it induces a decrease of this peroxidase in coleoptiles and an increase in mesocotyls. The peroxidase changes precede the growth changes. As measured by video stereomicroscopy or a position transducer, red light (R), which photoactivates phytochrome, stimulates coleoptile elongation with a lag of about 15-20 min and suppresses mesocotyl growth with a lag of 45-50 min. R also induces a 50% reduction in the extractable level of the anionic peroxidase in coleoptile walls in less than 10 min and a 40% increase in the level of this peroxidase in mesocotyl walls within 30 min. Ascorbic acid, an inhibitor of peroxidase activity, blocks the effects of R on mesocotyl section growth.These results are relevant to hypotheses that postulate that certain wall peroxidases can participate in light-induced changes in growth rate by their effects on wall extensibility.The regulatory pigment phytochrome can initiate rapid growth changes in plants. In etiolated seedlings of oats, maize, and other grasses, the photoactivation of phytochrome by red light (R) stimulates two distinct growth responses: it promotes coleoptile elongation and inhibits mesocotyl elongation (1).Changes in the extensibility of cell walls can help to mediate certain environmentally stimulated growth changes in plants (2), including those induced by phytochrome (3). Cell wall extensibility is at least partially controlled in many plants by one or more wall-localized enzymes that catalyze the formation or breakage of cell wall bonds (4). Consistent with this finding, several authors have noted that there is an inverse correlation between wall peroxidase activity and the growth of cell walls (5); e.g., when cucumber hypocotyl elongation is inhibited by blue light, there is an increase in wall peroxidase activity (6). The inverse correlation is consistent with the function of wall peroxidases in cross-linking wall macromolecules (7). To the extent peroxidases can decrease wall extensibility, they may help to mediate the inhibitory effects Ca2+ can have on wall extensibility (8), for Ca2+ can stimulate both the activity and secretion of wall peroxidases (9).There are numerous wall isoperoxidases, and there has been little or no previously published information correlating induced growth changes with changes in the content of any specific wall peroxidase isozyme. We recently characterized a monoclonal antibody, mWP3, which specifically crossreacts with a Mr 98,000 anionic peroxidase, which represents 15% of the total peroxidases present in soluble extracts of wall proteins from maize seedlings (10). Analysis by immunogold localization methods revealed t...

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“…These molecular mass values include: horseradish, 40 kD (26); potato, 47 kD (6); turnip, 33.4 kD (18); barley, 44 kD (25); and tobacco, 37 kD (12). An anionic molecular mass 98 kD wall peroxidase and a cationic molecular mass 58 kD wall peroxidase have been reported in maize seedlings (10). In addition, ethylene-inducible 33 kD (cationic) and 60 kD (anionic) peroxidases have been reported in cucumber cotyledons (1).…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of Isoperoxidases Elicited by Aspergillus flavus in Cotton Ovule Cultures

Mellon¹

1991

Plant Physiol.

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Two anionic isoperoxidases were isolated from media of Aspergillus flavus-inoculated cotton (Gossypium hirsutum L.) ovule cultures and purified about 150-fold to apparent homogeneity by treatment with Cell Debris Remover and ion exchange chromatography on Accell QMA medium. These isoperoxidases were present in noninoculated cotton ovule cultures at low levels. The major activity peak (B) represented 90% of the recovered peroxidase activity and was electrophoretically homogeneous. The minor activity peak (A) was about 95% pure. Isoelectric focusing analysis showed that B was greater than 95% pure with respect to other peroxidase isozymes, while the enzyme in A was about 90% isozymically pure. Each isoperoxidase displayed a molecular mass of 56 kilodaltons by interpolation from denaturing gel electrophoresis. The B isozyme displayed a molecular mass of 55 kilodaltons by gel filtration chromatography. The pH optima for the cotton ovule isoperoxidases were similar, 5.0 for isozyme A and 6.0 for isozyme B. The isoelectric points for isozymes A and B were 4.2 and 4.4, respectively. Eugenol, guaiacol, and 3,3',5,5'-tetramethylbenzidine were good electron donor substrates, whereas 4-aminoantipyrine was a poor substrate. The absorption spectrum of the material in B revealed a major peak at 400 nanometers and a minor peak at 280 nanometers. The molar extinction coefficient at 400 nanometers (pH 7.0) was calculated to be 1.07 x 105 per square centimeter per mole.Amino acid analysis of isozyme B confirmed the acidic nature of this protein and identified a number of similarities to the anionic peroxidases from tobacco and potato. This glycoprotein was found to contain 12 to 14% sugar (by weight), mainly in the form of galactose and mannose.

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Production and Characterization of Monoclonal Antibodies to Wall-Localized Peroxidases from Corn Seedlings

Cited by 29 publications

References 22 publications

Peroxidase Activity in the Leaf Elongation Zone of Tall Fescue

Peroxidase Activity in the Leaf Elongation Zone of Tall Fescue

Phytochrome induces changes in the immunodetectable level of a wall peroxidase that precede growth changes in maize seedlings

Purification and Characterization of Isoperoxidases Elicited by Aspergillus flavus in Cotton Ovule Cultures

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