| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
|
Plant Physiol. (1999) 120: 371-382 Isolation of Tobacco Isoperoxidases Accumulated in Cell-Suspension Culture Medium and Characterization of Activities Related to Cell Wall Metabolism1
Institut de Biologie Moléculaire des Plantes, Unité Propre de Recherche A0406, Centre National de la Recherche Scientifique, 12 rue du Général Zimmer, 67000 Strasbourg, France
All of the most important guaiacol-type peroxidase (POX) isoforms accumulated in the culture medium of BY-2 tobacco (Nicotiana tabacum L. cv Bright Yellow 2) cells have been isolated. Five basic and two acidic isoforms were found. The four major isoforms (B2, B3, P1, and P2), all strongly basic, have been purified to homogeneity and partially sequenced. B2 and B3 are new isoforms showing high homology to only one POX isolated so far. Amino acid sequencing and specific activities indicated that basic isoPOXs constitute two pairs of strictly related isoforms (P1/P2 and B2/B3). Their specific activities measured in the presence of different substrates, as monolignols and NAD(P)H, indicated possible specialized functions in cell wall metabolism. Only P-type POXs were able to oxidize indoleacetic acid. Variations in pH could play a regulatory role by changing the relative contribution of different isoforms to total POX activity. Apart from cell culture medium, polyclonal antibodies obtained against P1 and P2 detected P1 in roots and in lower parts of stems. Immunocytochemical labeling indicated that P-type POXs were expressed in stem phloem and in phloem and epidermal cells of roots.
Peroxidases belong to a large family of enzymes able to oxidize
several different substrates in the presence of
H2O2. In higher plants the
number of isoforms may be extremely high, up to 40 genes corresponding
to isoperoxidases for each plant, and several other isoforms can be
generated by posttranscriptional modifications (Van Engelen et al.,
1991 A peculiar feature of POXs is their apparent redundancy, i.e. several
isoforms can ensure the same reactions. At present, the available
information does not allow the prediction of the putative in vivo
function of any POX, given its amino acid sequence, or an understanding
of the reason for the high number of isoforms (Esnault and Chibbar,
1997 The difficulty of recognizing a relationship between POXs with
conserved sequences and specific physiological functions is mostly due
to the reactivities that isoPOXs show toward several potential
substrates, at least in in vitro conditions (Lagrimini et al., 1997 The accession numbers for the sequences reported in this article are
P81512 (B2) and P81513 (B3).
Plant, Cell, and Protoplast Material
Protein Extraction and Enzyme Assays Leaf, stem, root, cell, and callus crude extracts were obtained by grinding tissues with a mortar and pestle in the presence of 4 volumes of extraction buffer (50 mM K-phosphate, pH 7.5, 1 mM ascorbate, 1 mM EDTA, 1 mM DTT, 0.3% Triton X-100, and 0.8 M NaCl). Homogenate was filtered through cheesecloth and centrifuged for 30 min at 12,000g, and supernatants were recovered. Protoplast and derived-cell lysates were obtained by adding 3 volumes of extraction buffer to pelleted protoplasts or cells in Eppendorf tubes and by grinding the suspension with a minipestle. Spent media from protoplast and cell cultures were concentrated by freeze drying and dialyzed against 25 mM Tris-HCl, pH 7.5, buffer. Crude extracts and lysates were washed and concentrated using Centricon-10 tubes (Amicon) and the same buffer. Resulting samples were exploited for native- and SDS-PAGE, western blotting, and determination of control POX enzymic activity using o-dianisidine as a substrate, according to the method of Church and Galston (1988) .
Purified isoPOXs were used to calculate specific activities in the
presence of IAA, NAD(P)H, and monolignols. One unit of activity is
defined as the amount of enzyme that oxidizes 1 µmol of substrate per minute in standard conditions. NADH and NADPH POX-dependent oxidation were followed in the spectrophotometer by decrease in
A340 at pH 7.5 and 25°C, using 0.2 mM
H2O2 and 0.15 mM NAD(P)H, and an extinction coefficient of 6.22 mM 1
cm1 was used to calculate activity units.
Sinapyl and coniferyl alcohol oxidation were followed at
A272 and A264,
respectively, in the presence of 20 mM
K-phosphate buffer, pH 6.7, 0.3 mM monolignols, and 2 mM
H2O2, at 25°C; extinction
coefficient values of 8.09 and 13.09, respectively, were exploited
(Sterjiades et al., 1993). IAA oxidase activity was detected at
A261, according to Smith et al. (1982), by
using 50 mM Na-acetate buffer, pH 5.0, 0.2 mM IAA. pH-dependent isoPOX specific activities
were measured in the range of 4.2 to 7.7, using 50 mM citrate
Na2HPO4 (pH 4.2-5.2), 50 mM Na-acetate (pH 5.7-6.7), or 50 mM Tris-HCl (pH 7.2-7.7) buffers.
Purification of Isoperoxidases Columns and substrates used for protein purification (Q and SP Sepharose, Mono-Q, Mono-S, Phenyl-Superose, and Superdex 75) were from Pharmacia Biotech. Seven-day tobacco BY-2 cell-culture medium was recovered by filtration through nylon membrane and its volume reduced by freeze drying. Most of the pectic polysaccharides were removed by ultracentrifugation, and spent medium was dialyzed against ion-exchange chromatography buffer (25 mM Na-acetate, pH 5.2, or 25 mM Tris-HCl, pH 8.0, for SP and Q Sepharose columns, respectively) at 4°C, ultracentrifuged again to remove additional particulates, and loaded onto ion-exchange chromatography columns in the cold room. Alternatively, filtered cells were washed with water and resuspended in 0.5 M CaCl2. Ion-extracted proteins were recovered by filtration and processed as described above; no other isoPOXs were recovered, even though a different ratio among isoforms was observed. Moreover, no additional POX was found in NaCl-washed cells. Proteins were eluted with a NaCl linear gradient of 0 to 250 mM; fractions corresponding to successive peaks of POX activity were pooled separately, dialyzed overnight against the corresponding chromatography buffers, passed through a 0.22 µM protein filter (Millipore), and further purified using fast-protein liquid chromatography at room temperature. Samples separated by SP and Q Sepharose columns were loaded onto Mono-S and Mono-Q columns, respectively, and eluted with linear gradients of different NaCl concentrations (from 0 to 100 or 200 mM) to maximize peak separation.
IEF-, Native-, and SDS-PAGE Ampholine carrier ampholytes, an IEF calibration kit, and SDS-PAGE molecular-mass standards were purchased from Pharmacia Biotech Europe. IEF-PAGE was run as described by Robertson et al. (1987) over a pH
range of 3.5 to 10.0. Native- and SDS-PAGE (10% when not indicated
otherwise) were run according to the method of Laemmli (1970). SDS gels
were silver stained (Eschenbruck and Bürch, 1982), and POXs were
visualized in native and IEF gels using chloro-1-naphthol (Lagrimini
and Rothstein, 1987). Glycoproteins were stained by immersion of gels
in Schiff's reagent (Merck, Darmstadt, Germany) after periodic
oxidation, according to the manufacturer's instructions.
Western Blotting SDS-PAGE was run in standard conditions, and gels were removed, equilibrated in transfer buffer (25 mM Tris-HCl, pH 8.0, 192 mM Gly, and 15% methanol) for 10 min, and mounted in a transblotting sandwich using an Immobilon-P membrane (Millipore). After electrotransfer (45 min at 400 mA), membranes were stained with Ponceau S solution (Serva, Heidelberg, Germany) for protein detection. Immunodetection was performed according to the method of Geoffroy et al. (1990).
Protein Microsequencing Purified proteins were run in standard conditions on denaturing gels, equilibrated in 10 mM 3-cyclohexylamino-1-propanesulfonic acid, pH 11.0, 10% methanol buffer, and electroblotted onto a Problott membrane (Perkin-Elmer Applied Biosystems, Courtaboeuf, France) for 30 min at 50 V, at room temperature. The membrane was washed and stained with Coomassie Blue R-250 (Merck), and protein bands were recovered for N-terminal sequencing, according to the method of Heitz et al. (1994). Partial
proteolysis was used to sequence N-terminal blocked proteins. Purified
proteins (30-50 µg) were run on denaturing gels and stained in
Coomassie Blue R-250, and bands corresponding to the proteins were cut, soaked in 100 mM Tris-HCl, pH 6.8, 1 mM EDTA,
and 0.1% SDS, and loaded into a well of a polyacrylamide denaturing
gel. Into the same well was deposited 2 to 5 µg of endoproteinase
Glu-C (Boehringer Mannheim) dissolved in the equilibration buffer plus
10% glycerol. The migration front was allowed to reach the interface
between stacking and running (15%) gels by using standard PAGE
conditions before the power was switched off. After 30 min, the
products of the proteolysis reaction were separated in the running gel and transferred onto a Problott membrane, as described above. Microsequencing of peptides was performed as described by Geoffroy et
al. (1990).
Antibody Preparation A P1/P2 isoPOX mixture was dialyzed against 0.9% NaCl. Two-hundred micrograms of protein was dissolved in a final volume of 300 µL and combined with an equal volume of Freund's complete adjuvant, and the emulsion was injected subcutaneously into a rabbit. Eighty micrograms of protein was combined with Freund's incomplete adjuvant and injected intramuscularly three times (every 2 weeks for 6 weeks) to booster the antibody reaction. IgG was purified from sera by chromatography, exploiting a Fractogel EMD TA column (Merck), and stored with 20% glycerol at 20°C.
Immunocytolocalization Tissues were fixed and subsequently embedded in paraffin and 10-µm-thick cuts were performed. After deparaffination and dehydration, samples were washed in PBS in the presence of 0.05% Triton X-100 and 1% BSA. Surfaces were saturated using the same buffer plus 5% goat serum (Boehringer Mannheim) and incubated at 4°C overnight in PBS, 0.01% Triton X-100, 1% BSA, and primary antibodies diluted 1:1000. Samples were washed in the same buffer, incubated for 1 h at room temperature in the presence of anti-rabbit alkaline phosphatase-conjugated secondary antibodies (1:1000 in PBS; Boehringer Mannheim), washed in PBS, and equilibrated with 100 mM Tris-HCl, pH 8.2. Color reaction was developed using Fast Red (Sigma) and blocked by 10 mM Tris-HCl, pH 8.0, 1 mM EDTA. Finally, samples were washed in water and mounted in glycerol. Controls had no antibodies or preimmune serum added instead of primary antibodies. Callose in cell walls of phloem cells was visualized by fluorescence with decolorized aniline blue according to the method of Currier and Strugger (1956).
Other Methods Protein concentration was determined by the Bio-Rad protein assay kit using BSA as a protein standard. Sequence data were analyzed using the FASTA search service (Pearson and Lipman, 1988) and the Genetics
Computer Group (Madison, WI) package programs (Devereux et al., 1984).
Purification of Tobacco IsoPOXs We chose to work with isoforms accumulated in BY-2 cell-suspension culture medium and to study their role in cell wall metabolism. Preliminary tests established that 7-d-old cultures gave the highest yield of POXs and that POX specific activity increased 4-fold between d 2 and 7 of cell culture, even though the pattern of expression of the isoforms did not change (data not shown). The standard protocol of purification for the different isoPOXs is described in Figure 1. Two steps of centrifugation, before and after dialysis against ion-exchange chromatography buffer, were necessary to eliminate most of the abundant polysaccharides present in the filtrate that resulted from the freeze-drying concentration of the medium. About 95% of POX activity preferentially bound to a cation exchanger (SP Sepharose column), whereas the recovery of Q Sepharose-bound fractions (anion exchanger) was very low, regardless of the protocol (standard or alternative) used for their purification (see ``Materials and Methods''). This indicates that very basic isoforms were predominant in the cell-suspension culture medium (Table I). The two most important acidic isoforms, A1 and A2, were purified according to the protocol described in Figure 1. They had pI values of 3.9 (A1) and 4.5 (A2), respectively (Fig. 2A), and very different mobility in native gel (Fig. 2B). Even though A2 showed stronger activity than A1 using chloro-1-naphthol in native gels loaded with filtered medium (Fig. 2A), the scarce amount obtained after the Mono-Q elution step prevented any further purification. In contrast, enough A1 was available to purify an isoform of 45 kD to homogeneity (Table II; Fig. 3A).
Microsequencing of IsoPOXs B2, B3, P1, and P2 B2, B3, and A1 were N blocked; this feature is quite frequent in POXs, due to a cyclization of a Gln in position 1 to pyroglutamate (Kjærsgård et al., 1997). B2 and B3 were partially digested to obtain
peptides that could be sequenced. Sequences of 83 and 60 amino acids
located at their N and C termini, respectively, were determined for
each isoPOX, which represent almost one-half of the length of a common
POX (Fig. 4). The two isoforms were very similar; they differed in only 3 amino acids over the available sequence. B2 and B3 have not been described and were very close to a
POX isolated from peanut cell-culture medium (Buffard et al., 1990):
68% identity and 84% similarity in the N-terminal region; 77%
identity and 92% similarity in the C-terminal region (Fig. 4). B2 and
B3 showed less identity to other basic POXs, such as wheat POX, P1/P2,
Arabidopsis, and horseradish POXs (about 50% identity and 70%
similarity in the N-terminal region; about 40% identity and between
51% and 59% similarity in the C-terminal region). P1 and P2 were
N-terminal microsequenced for 48 and 42 amino acids, respectively. They
showed perfect homology to the predicted amino acid sequences
corresponding to the previously reported clones D42064 and D42065
(Narita et al., 1995), except that they contained an additional Val in
position 23 of the mature protein that was not reported by Narita et
al. (1995).
pH Dependence of IsoPOX Activity The activities of the four major basic isoPOXs (B2, B3, P1, and P2) and that of A1 were measured while varying the pH of the buffer in the reaction mixture from 4.2 to 7.7. B2/B3 isoPOXs and A1 had a maximum of activity at pH 4.7, and A1 conserved a low activity at slightly alkaline pH values (Fig. 5). P1/P2 showed a wider range of suitable pH values than B2/B3 and A1 POXs, with a maximum of activity at pH 5.2.
Substrate Specificity of IsoPOXs in Vitro Three types of substrates were tested: (a) NADP and NADPH, because POXs can trigger the nonenzymic production of H2O2 in the presence of NAD(P)H, which is later used as a substrate for the oxidation of phenolic groups (Halliwell, 1978); (b) monolignols, such as coniferyl
and sinapyl alcohol, which are polymerized in secondary cell walls
(Eberhardt et al., 1993); and (c) IAA, which POXs may catabolize in
vivo (Gazaryan et al., 1996). The activity of B1 was barely detectable
and then only in the presence of monolignols (Table
III). The other purified POXs could use
both NAD(P)H and monolignols as the substrates. A1 had the highest
specific activity with both NAD(P)H and monolignols compared with B-
and P-type isoPOXs (Table III). A fraction enriched in A2 and
containing negligible contamination due to other POXs (Fig. 2) showed
relatively low affinity for NAD(P)H, but a higher specific activity
with monolignols than other isoPOXs (data not shown). P-type isoPOXs
showed higher affinity for NADPH than B-type isoPOXs, which, in
contrast, could use monolignols with more elevated efficiency than
P-type isoPOXs (Table III). Furthermore, P-type isoPOXs were the only
ones that possessed IAA oxidase activity (Table III).
Immunodetection Using Polyclonal Antibodies against P1 and P2 A mixture of the strictly related isoPOXs P1 and P2 was used as antigen for the production of polyclonal antibodies. Protein extracts from different plant organs and from BY2 cells were analyzed by western blotting. Specific signals corresponding to P1 could be detected in the lower parts of stems, both nodes and internodes, as well as in roots (Fig. 6, lanes LN, LI, and R). No signal could be detected in leaves and in the upper parts of stems (Fig. 6, lanes YL, ML, UN, and UI). Finally, weak signals corresponding to P1 and P2 appeared in the cell extract, whereas very strong signals were detected in the spent medium (Fig. 6, lanes C and M). It should be noted that the amount of protein loaded in lane M was only one-tenth of that loaded in the other lanes. P2 was also detected in extracts from BY2 calli grown on solid medium, whereas only a faint signal corresponding to P1 appeared (data not shown). The other weak signals detected in the different samples may correspond to other POXs sharing common epitopes with P-type isoPOXs. Regenerating protoplasts isolated from tobacco leaves were used to determine if P-type isoPOXs accumulated at specific times during culture. Separate aliquots of both protoplasts and their corresponding media were recovered daily during the 1st week of culture and used for western blotting, but no protein was recognized by anti-P-type antibodies (data not shown). In conclusion, apart from BY2 cells and culture medium, the presence of the P1 protein was detected in roots and in stems, whereas the P2 protein could not be found.
Cell wall metabolism is very important for protoplast and cell
development, and POXs are considered to play a primary role in
polymerizing structural elements and regulating cell wall elasticity during cell elongation (Hoson et al., 1995 Received December 21, 1998;
accepted March 15, 1999.
Abbreviation:
POX, guaiacol-type peroxidase.
The authors thank Dr. Roberte Bronner for her assistance in
microscopy, Pierrette Geoffroy for her collaboration with fast-protein liquid chromatography, and Monique Le Ret for microsequencing of
proteins.
Asada K
(1992)
Ascorbate peroxidase: a hydrogen peroxide-scavenging enzyme in plants.
Physiol Plant
85:
235-241
[CrossRef]
Botella MA,
Quesada MA,
Kononowicz AK,
Bressan RA,
Pliego F,
Hasagawa PM,
Valpuesta V
(1994)
Characterization and in situ localization of a salt-induced tomato peroxidase mRNA.
Plant Mol Biol
25:
105-114
[CrossRef][Web of Science][Medline]
Boyer N, De Jaegher G (1986) Direct or indirect role of
peroxidases in ethylene biosynthesis? In H Greppin, C Penel,
T Gaspar, eds, Molecular and Physiological Aspects of Plant
Peroxidases. University of Geneva Press, Geneva, pp 47-60
Bradley DJ,
Kjellbom P,
Lamb CJ
(1992)
Elicitor- and wound-induced oxidative cross-linking of a plant cell wall proline-rich protein: a novel, rapid defense response.
Cell
70:
21-30
[CrossRef][Web of Science][Medline]
Breda C,
Buffard D,
van Huystee RB,
Esnault R
(1993)
Differential expression of two peanut peroxidase cDNA clones in peanut plants and cells in suspension culture in response to stress.
Plant Cell Rep
12:
268-272
Brisson LF,
Tenhaken R,
Lamb C
(1994)
Function of oxidative cross-linking of cell wall structural proteins in plant disease resistance.
Plant Cell
6:
1703-1712
Brownleader MD,
Ahmed N,
Trevan M,
Chaplin MF,
Dey PM
(1995)
Purification and partial characterization of tomato extensin peroxidase.
Plant Physiol
109:
1115-1123
[Abstract]
Buffard D,
Breda C,
van Huystee RB,
Asemota O,
Pierre M,
Dang Ha DB,
Esnault R
(1990)
Molecular cloning of complementary DNAs encoding two cationic peroxidases from cultivated peanut cells.
Proc Natl Acad Sci USA
87:
8874-8878
Chen G-H,
Asada K
(1989)
Ascorbate peroxidase in tea leaves: occurrence of two isozymes and their differences in enzymatic and molecular properties.
Plant Cell Physiol
30:
987-998
Church DL,
Galston AW
(1988)
4-Coumarate coenzyme A ligase and isoperoxidase expression in Zinnia mesophyll cells induced to differentiate into tracheary elements.
Plant Physiol
88:
679-684
Cooper JB,
Heuser JE,
Varner JE
(1994)
3,4-Dehydroproline inhibits cell wall assembly and cell division in tobacco protoplasts.
Plant Physiol
104:
747-754
[Abstract]
Criqui M-C,
Plesse B,
Durr A,
Marbach J,
Parmentier Y,
Jamet E,
Fleck J
(1992)
Characterization of genes expressed in mesophyll protoplasts of Nicotiana sylvestris before the re-initiation of the DNA replicational activity.
Mech Dev
38:
121-132
[Medline]
Currier H,
Strugger S
(1956)
Aniline blue and fluorescence microscopy of callose in bulbscales of Allium cepa L.
Protoplasma
45:
552-559
[CrossRef][Web of Science]
de Marco A,
Roubelakis-Angelakis KA
(1996a)
The complexity of enzymic control of hydrogen peroxide may affect the regeneration potential of plant protoplasts.
Plant Physiol
110:
137-145
[Abstract]
de Marco A,
Roubelakis-Angelakis KA
(1996b)
Hydrogen peroxide plays a bivalent role in the regeneration of protoplasts.
J Plant Physiol
149:
109-114
de Marco A,
Roubelakis-Angelakis KA
(1997)
Laccase activity could contribute to cell-wall reconstitution in regenerating protoplasts.
Phytochemistry
46:
421-425
[CrossRef]
Devereux J,
Haeberli P,
Smithies O
(1984)
A comprehensive set of sequence analysis programs for the VAX.
Nucleic Acids Res
12:
387-395
Eberhardt TL,
Bernards MA,
He L,
Davin LB,
Wooten JB,
Lewis NG
(1993)
Lignification in cell suspension cultures of Pinus taeda.
J Biol Chem
28:
21088-21096
Eschenbruck M,
Bürch RM
(1982)
Experimentally improved relationship of ultrasensitive silver staining of protein in polyacrylamide gels.
Anal Biochem
125:
96-99
[CrossRef][Web of Science][Medline]
Eshdat Y,
Holland D,
Faltin Z,
Ben-Hayyim G
(1997)
Plant glutathione peroxidases.
Physiol Plant
100:
234-240
[CrossRef]
Esnault R,
Chibbar RN
(1997)
Peroxidases: at the gene expression level.
Plant Peroxidase Newsletter
10:
7-14
Everdeen DS,
Kiefer S,
Willard JJ,
Muldoon EP,
Dey PM,
Li X-B,
Lamport DTA
(1988)
Enzymic crosslinkage of monomeric extensin precursors in vitro.
Plant Physiol
87:
616-621
Fujiyama K,
Takemura H,
Shibayama S,
Kobayashi K,
Choi JK,
Shinmio A,
Takano M,
Yamada Y,
Okada M
(1988)
Structure of the horseradish peroxidase isozyme C genes.
Eur J Biochem
73:
681-687
Gazaryan IG,
Lagrimini LM,
Ashby GA,
Thorneley R
(1996)
The mechanism of indole-3-acetic acid oxidation by plant peroxidases: anaerobic stopped-flow spectrophotometric studies on horseradish and tobacco peroxidases.
Biochem J
313:
841-847
Geoffroy P,
Legrand M,
Fritig B
(1990)
Isolation and characterization of a proteinaceous inhibitor of microbial proteinases induced during the hypersensitive reaction of tobacco to tobacco mosaic virus.
Mol Plant Microbe Interact
3:
327-333
[Web of Science][Medline]
Goodspeed TH
(1954)
Origins, relationships and evolution of its species in the light of their distribution, morphology and cytogenetics.
In
TH Goodspeed,
eds, The Genus Nicotiana. Chronica Botanica
Acta, Waltham, MA, pp 357-391
Gross GG,
Janse C,
Elstner EF
(1977)
Involvement of malate, monophenols and the superoxide radical in hydrogen peroxide formation by isolated cell wall from horseradish (Armoracia lapathifolia Gilib).
Planta
136:
271-276
[CrossRef]
Halliwell B
(1978)
Lignin synthesis: the generation of hydrogen peroxide and superoxide by peroxidase and its stimulation by manganese (II) and phenols.
Planta
140:
81-88
[CrossRef]
Heitz T,
Segond S,
Kauffmann S,
Geoffroy P,
Prasad V,
Brunner F,
Fritig B,
Legrand M
(1994)
Molecular characterization of a novel tobacco pathogenesis-related (PR) protein: a new plant chitinase/lysozyme.
Mol Gen Genet
245:
246-254
[Web of Science][Medline]
Hertig C,
Rebmann G,
Bull J,
Mauch F,
Dudler R
(1991)
Sequence and tissue-specific expression of a putative peroxidase gene from wheat (Triticum aestivum L.).
Plant Mol Biol
16:
171-174
[Medline]
Hoson T,
Wakabayashi K,
Masuda Y
(1995)
Inhibition of the breakdown of xyloglucans in azuki bean epicotyls by concanavalin A.
Plant Cell Physiol
36:
897-902
Intapruk C,
Higashimura N,
Yamamoto K,
Okada N,
Shinmyo A,
Takano M
(1991)
Nucleotide sequences of two genomic DNAs encoding peroxidases of Arabidopsis thaliana.
Gene
98:
237-241
[Medline]
Intapruk C,
Yamamoto K,
Sekine M,
Takano M,
Shinmyo A
(1994)
Regulatory sequences involved in the peroxidase gene expression in Arabidopsis thaliana.
Plant Cell Rep
13:
123-129
Keller,
CP,
Van Volkenburgh E
(1996)
Osmoregulation by oat coleoptile protoplasts.
Plant Physiol
110:
1007-1016
[Abstract]
Kjærsgård IVH,
Jespersen HM,
Rasmussen SK,
Welinder KG
(1997)
Sequence and RT-PCR expression analysis of two peroxidases from Arabidopsis thaliana belonging to a novel evolutionary branch of plant peroxidases.
Plant Mol Biol
33:
699-708
[CrossRef][Web of Science][Medline]
Klotz KL,
Liu T-TY,
Liu L,
Lagrimini LM
(1998)
Expression of the tobacco anionic peroxidase gene is tissue-specific and developmentally regulated.
Plant Mol Biol
36:
509-520
[CrossRef][Web of Science][Medline]
Koop HV,
Schweiger HG
(1985)
Regeneration of plants from individually cultivated protoplasts using an improved microculture system.
J Plant Physiol
121:
245-257
Laemmli UK
(1970)
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227:
680-685
[CrossRef][Medline]
Lagrimini LM
(1991)
Wound-induced deposition of polyphenols in transgenic plants overexpressing peroxidase.
Plant Physiol
96:
577-583
Lagrimini LM,
Gingas V,
Finger F,
Rothstein S,
Liu T-TY
(1997)
Characterization of antisense transformed plants deficient in the tobacco anionic peroxidase.
Plant Physiol
114:
1187-1196
[Abstract]
Lagrimini LM,
Rothstein S
(1987)
Tissue specificity of tobacco peroxidase isozymes and their induction by wounding and tobacco mosaic virus infection.
Plant Physiol
84:
438-442
Marañon MJR,
van Huystee RB
(1994)
Plant peroxidases: interaction between their prosthetic groups.
Phytochemistry
37:
1217-1225
[CrossRef]
Melo NS,
Larsen E,
Welinder KG,
Fevereiro PS
(1997)
Characterization of two major cationic peroxidases from cell suspension cultures of Vaccinum myrtillus.
Plant Sci
122:
1-10
[CrossRef]
Mohan R,
Vijayan P,
Kolattukudy PE
(1993)
Developmental and tissue-specific expression of a tomato anionic peroxidase (tap1) gene by a minimal promoter, with wound and pathogen induction by an additional 5
Nagata T,
Nemoto Y,
Kasezawa S
(1992)
Tobacco BY2 cell line as the "Hela" cell in the cell biology of higher plants.
Int Rev Cytol
132:
1-30
[Web of Science]
Narita H,
Asaka Y,
Ikura K,
Matsumoto S,
Sasaki R
(1995)
Isolation, characterization and expression of cationic peroxidase isozymes released into the medium of cultured tobacco cells.
Eur J Biochem
228:
855-862
[Medline]
Østergaard L,
Abelskov AK,
Mattsson O,
Welinder KG
(1996)
Structure and organ specificity of an anionic peroxidase from Arabidopsis thaliana cell suspension culture.
FEBS Lett
398:
243-247
[CrossRef][Web of Science][Medline]
Pearson WR,
Lipman DJ
(1988)
Improved tools for biological sequence comparison.
Proc Natl Acad Sci USA
85:
2444-2448
Robertson EF,
Dannelly HK,
Malloy PJ,
Reeves HC
(1987)
Rapid isoelectric focusing in a vertical polyacrylamide minigel system.
Anal Biochem
167:
290-294
[CrossRef][Web of Science][Medline]
Schnabelrauch LS,
Kieliszewski M,
Upham BL,
Alizedeh H,
Lamport DTA
(1996)
Isolation of pI 4.6 extensin peroxidase from tomato cell suspension cultures and identification of Val-Tyr-Lys as putative intermolecular cross-link site.
Plant J
9:
477-489
[CrossRef][Web of Science][Medline]
Scott-Craig JS,
Kerby K,
Stein BD,
Sommerville SC
(1995)
Expression of an extracellular peroxidase that is induced in barley (Hordeum vulgare) by the powdery mildew pathogen (Erysiphe graminis).
Physiol Mol Plant Pathol
47:
407-418
[CrossRef]
Sherf BA,
Bajar AM,
Kolattukudy PE
(1993)
Abolition of an inducible highly anionic peroxidase activity in transgenic tomato.
Plant Physiol
101:
201-208
[Abstract]
Smith AM,
Morrison WL,
Milham PJ
(1982)
Oxidation of indole-3-acetic by peroxidase: involvement of reduced peroxidase and compound III with superoxide as a product.
Biochemistry
21:
4414-4419
[Medline]
Sterjiades R,
Dean JFD,
Gamble G,
Himmelsbach DS,
Erikson K-EL
(1993)
Extracellular laccase and peroxidases from sycamore maple (Acer pseudoplatanus) cell suspension cultures: reactions with monolignols and lignin model compounds.
Planta
190:
75-87
Thordal-Christensen H,
Brandt J,
Cho BH,
Rasmussen SK,
Gregersen PL,
Smedegaard-Petersen V,
Collinge DB
(1992)
cDNA cloning and characterization of two barley peroxidase transcripts induced differentially by the powdery mildew fungus Erysiphe graminis.
Physiol Mol Plant Pathol
40:
395-409
[CrossRef]
Van Engelen FA,
Sterk P,
Booij H,
Cordewener JHG,
Rook W,
van Kammen A,
deVries SC
(1991)
Heterogeneity and cell type-specific localization of a cell wall glycoprotein from carrot suspension cells.
Plant Physiol
96:
705-712
Welinder KG
(1985)
Plant peroxidases: their primary, secondary and tertiary structures, and relation to cytochrome c peroxidase.
Eur J Biochem
151:
497-504
[Medline]
Welinder KG, Jespersen HN, Kjærsgård IVH, Østergaard L, Abelskov AK,
Hansen LN, Rasmussen SK (1996) What can we learn from
Arabidopsis peroxidases? In C Obinger, U Burner,
R Ebermann, C Penel, H Greppin, eds, Plant Peroxidases: Biochemistry
and Physiology. University of Geneva Press, Geneva, pp
173-178
This article has been cited by other articles:
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| ASPB Publications | PLANT PHYSIOLOGY® | THE PLANT CELL | |
|---|---|---|---|
Top
Abstract
Introduction
), B-type (
),
and P-type (
) POXs. Specific activities were measured as described
in ``
-flanking region.
Plant Mol Biol
22:
475-490
