Abstract
Membranes from tobacco cell suspension cultures were used as antigens for the preparation of monoclonal antibodies. Use of solid phase and indirect immunofluorescence assays led to the identification of hybridomas producing antibodies directed against cell surface epitopes. One of these monoclonal antibodies (11.D2) was found to recognize a molecular species which on two-dimensional analysis (using nonequilibrium pH-gradient electrophoresis and SDS-PAGE) was found to have a high and polydisperse molecular mass and a very basic isoelectric point. This component was conspicuously labeled by [3H]proline in vivo. The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan. Use of the monoclonal antibody as an immunoaffinity reagent allowed the purification of a tobacco glycoprotein which was identical in amino acid composition to extensin. Finally, immunocytological analyses revealed tissue-specific patterns of labeling by the monoclonal antibody that were identical to those observed with a polyclonal antibody raised against purified extensin. We have concluded that monoclonal antibody 11.D2 recognizes an epitope that is carried exclusively by extensin. Analysis of cellular homogenates through differential and isopycnic gradient centrifugation revealed that biosynthesis of the extensin epitope was found on or within the membranes of the endoplasmic reticulum, Golgi region and plasma membrane. This result is consistent with the progressive glycosylation of the newly-synthesized extensin polypeptide during its passage through a typical eukaryotic endomembrane pathway of secretion. The 11.D2 epitope was not found in protoplasts freshly isolated from leaf tissues. However, on incubation of these protoplasts in appropriate culture media, biosynthesis of the epitope was initiated. This process was not impeded by the presence of chemicals that are reported to be inhibitors of cell wall production or of proline hydroxylation.
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Claims 13
Claim · model-written summary
Assays identified antibody-producing hybridomas.
- Taxon
- tobacco
Nicotiana tabacumt0 - Outcome
- identification of hybridomase0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Use of solid phase and indirect immunofluorescence assays led to the identification of hybridomas producing antibodies directed against cell surface epitopes.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Antibody 11.D2 recognized a high-mass basic molecular species.
- Taxon
- tobaccoNicotiana tabacumt0
- Gene
- 11.D2e0
- Outcome
- high and polydisperse molecular mass and a very basic isoelectric pointe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“One of these monoclonal antibodies (11.D2) was found to recognize a molecular species which on two-dimensional analysis (using nonequilibrium pH-gradient electrophoresis and SDS-PAGE) was found to have a high and polydisperse molecular mass and a very basic isoelectric point.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
The component was labeled by proline.
- Taxon
- tobaccoNicotiana tabacumt0
- Outcome
- labeled by [3H]proline in vivoe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This component was conspicuously labeled by [3H]proline in vivo.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
The antibody reacted with tomato extensin.
- Taxon
- tomato
Solanum lycopersicume0 - Outcome
- cross-reacted with authentic tomato extensine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan.”
Claim · model-written summary
It did not react with potato lectin.
- Taxon
- potato
Solanum tuberosume0 - Outcome
- not with potato lectine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan.”
Claim · model-written summary
It did not react with larch arabinogalactan.
Verbatim
e0“The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan.”
Claim · model-written summary
Immunoaffinity purification yielded tobacco glycoprotein matching extensin.
- Taxon
- tobaccoNicotiana tabacume0
- Outcome
- purification of a tobacco glycoproteine0
- Comparator
- extensine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Use of the monoclonal antibody as an immunoaffinity reagent allowed the purification of a tobacco glycoprotein which was identical in amino acid composition to extensin.”
Claim · model-written summary
Monoclonal and polyclonal antibodies gave identical tissue labeling.
- Taxon
- tobaccoNicotiana tabacumt0
- Outcome
- tissue-specific patterns of labelinge0
- Comparator
- polyclonal antibodye0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“immunocytological analyses revealed tissue-specific patterns of labeling by the monoclonal antibody that were identical to those observed with a polyclonal antibody raised against purified extensin.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Antibody 11.D2 recognizes an epitope exclusive to extensin.
- Taxon
- tobaccoNicotiana tabacumt0
- Gene
- extensine0
- Outcome
- epitope that is carried exclusively by extensine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“monoclonal antibody 11.D2 recognizes an epitope that is carried exclusively by extensin.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Extensin epitope synthesis occurred at three membrane compartments.
- Taxon
- tobaccoNicotiana tabacumt0
- Organ / stage
- endoplasmic reticulum, Golgi region and plasma membranee0
- Gene
- extensine0
- Outcome
- biosynthesis of the extensin epitopee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“biosynthesis of the extensin epitope was found on or within the membranes of the endoplasmic reticulum, Golgi region and plasma membrane.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Fresh leaf protoplasts lacked the epitope.
- Taxon
- tobaccoNicotiana tabacumt0
- Organ / stage
- protoplastse0
- Outcome
- 11.D2 epitope was not founde0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The 11.D2 epitope was not found in protoplasts freshly isolated from leaf tissues.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Culture incubation initiated epitope biosynthesis.
- Taxon
- tobaccoNicotiana tabacumt0
- Treatment
- incubation of these protoplasts in appropriate culture mediae0
- Outcome
- biosynthesis of the epitopee0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“on incubation of these protoplasts in appropriate culture media, biosynthesis of the epitope was initiated.”
t0“tobacco cell suspension cultures”
Claim · model-written summary
Inhibitory chemicals did not impede epitope biosynthesis.
- Taxon
- tobaccoNicotiana tabacumt0
- Treatment
- chemicals that are reported to be inhibitors of cell wall production or of proline hydroxylatione0
- Outcome
- This processe0
- Direction
- not impedede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This process was not impeded by the presence of chemicals that are reported to be inhibitors of cell wall production or of proline hydroxylation.”
t0“tobacco cell suspension cultures”
The same questions in other papers
in vitro · molecular: metabolites · Nicotiana tabacum (tobacco)Positive and negative results
9 papers in the same direction as this paper · 6 in the opposite direction
- negative
- negative
- no effect
12 papers more in this group
See all 16 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.