Plant-based heterologous expression of Mal d 2, a thaumatin-like protein and allergen of apple (Malus domestica), and its characterization as an antifungal protein.
Identifieur interne : 000471 ( Main/Corpus ); précédent : 000470; suivant : 000472Plant-based heterologous expression of Mal d 2, a thaumatin-like protein and allergen of apple (Malus domestica), and its characterization as an antifungal protein.
Auteurs : Monika Krebitz ; Birgit Wagner ; Fatima Ferreira ; Clemens Peterbauer ; Nuria Campillo ; Michael Witty ; Daniel Kolarich ; Herta Steinkellner ; Otto Scheiner ; Heimo BreitenederSource :
- Journal of molecular biology [ 0022-2836 ] ; 2003.
English descriptors
- KwdEn :
- Allergens (chemistry), Allergens (genetics), Allergens (pharmacology), Antifungal Agents (chemistry), Antifungal Agents (pharmacology), Cloning, Molecular (MeSH), DNA, Complementary (MeSH), Food Hypersensitivity (blood), Food Hypersensitivity (etiology), Food Hypersensitivity (immunology), Fusarium (drug effects), Gene Expression Regulation, Plant (MeSH), Humans (MeSH), Immunoblotting (MeSH), Immunoglobulin E (metabolism), Malus (MeSH), Models, Molecular (MeSH), Molecular Sequence Data (MeSH), Penicillium (drug effects), Peptide Fragments (metabolism), Plant Proteins (chemistry), Plant Proteins (genetics), Plant Proteins (pharmacology), Recombinant Proteins (MeSH), Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (MeSH), Sweetening Agents (chemistry), Sweetening Agents (pharmacology), Tobacco (chemistry), Tobacco Mosaic Virus (genetics), Transfection (MeSH).
- MESH :
- chemical , chemistry : Allergens, Antifungal Agents, Plant Proteins, Sweetening Agents.
- chemical , genetics : Allergens, Plant Proteins.
- chemical , metabolism : Immunoglobulin E, Peptide Fragments.
- chemical , pharmacology : Allergens, Antifungal Agents, Plant Proteins, Sweetening Agents.
- blood : Food Hypersensitivity.
- chemistry : Tobacco.
- drug effects : Fusarium, Penicillium.
- etiology : Food Hypersensitivity.
- genetics : Tobacco Mosaic Virus.
- immunology : Food Hypersensitivity.
- Cloning, Molecular, DNA, Complementary, Gene Expression Regulation, Plant, Humans, Immunoblotting, Malus, Models, Molecular, Molecular Sequence Data, Recombinant Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Transfection.
Abstract
Mal d 2 is a thaumatin-like protein and important allergen of apple fruits that is associated with IgE-mediated symptoms in apple allergic individuals. We obtained a full-length cDNA clone of Mal d 2 from RNA isolated from ripe apple (Malus domestica cv. Golden Delicious). The cDNA's open reading frame encodes a protein of 246 amino acid residues including a signal peptide of 24 residues and two putative glycosylation sites. The deduced amino acid sequence of the mature Mal d 2 protein results in a predicted molecular mass of 23,210.9Da and a calculated pI of 4.55. Sequence comparisons and molecular modeling place Mal d 2 among those pathogenesis-related thaumatin-like proteins that contain a conserved acidic cleft. In order to ensure the correct formation of the protein's eight conserved disulfide bridges we expressed Mal d 2 in Nicotiana benthamiana plants by the use of a tobacco mosaic viral vector. Transfected N.benthamiana plants accumulated Mal d 2 to levels of at least 2% of total soluble protein. MALDI-TOF mass spectrometric analyses of the recombinant Mal d 2 and its proteolytic fragments showed that the apple-specific leader peptide was correctly cleaved off by the host plant and that the mature recombinant protein was intact and not glycosylated. Purified recombinant Mal d 2 displayed the ability to bind IgE from apple-allergic individuals equivalent to natural Mal d 2. In addition, the recombinant thaumatin-like Mal d 2 exhibited antifungal activity against Fusarium oxysporum and Penicillium expansum, implying a function in plant defense against fungal pathogens.
DOI: 10.1016/s0022-2836(03)00403-0
PubMed: 12787673
Links to Exploration step
pubmed:12787673Le document en format XML
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Peterbauer</name></author><author><name sortKey="Campillo, Nuria" sort="Campillo, Nuria" uniqKey="Campillo N" first="Nuria" last="Campillo">Nuria Campillo</name></author><author><name sortKey="Witty, Michael" sort="Witty, Michael" uniqKey="Witty M" first="Michael" last="Witty">Michael Witty</name></author><author><name sortKey="Kolarich, Daniel" sort="Kolarich, Daniel" uniqKey="Kolarich D" first="Daniel" last="Kolarich">Daniel Kolarich</name></author><author><name sortKey="Steinkellner, Herta" sort="Steinkellner, Herta" uniqKey="Steinkellner H" first="Herta" last="Steinkellner">Herta Steinkellner</name></author><author><name sortKey="Scheiner, Otto" sort="Scheiner, Otto" uniqKey="Scheiner O" first="Otto" last="Scheiner">Otto Scheiner</name></author><author><name sortKey="Breiteneder, Heimo" sort="Breiteneder, Heimo" uniqKey="Breiteneder H" first="Heimo" last="Breiteneder">Heimo Breiteneder</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date 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Wagner</name></author><author><name sortKey="Ferreira, Fatima" sort="Ferreira, Fatima" uniqKey="Ferreira F" first="Fatima" last="Ferreira">Fatima Ferreira</name></author><author><name sortKey="Peterbauer, Clemens" sort="Peterbauer, Clemens" uniqKey="Peterbauer C" first="Clemens" last="Peterbauer">Clemens Peterbauer</name></author><author><name sortKey="Campillo, Nuria" sort="Campillo, Nuria" uniqKey="Campillo N" first="Nuria" last="Campillo">Nuria Campillo</name></author><author><name sortKey="Witty, Michael" sort="Witty, Michael" uniqKey="Witty M" first="Michael" last="Witty">Michael Witty</name></author><author><name sortKey="Kolarich, Daniel" sort="Kolarich, Daniel" uniqKey="Kolarich D" first="Daniel" last="Kolarich">Daniel Kolarich</name></author><author><name sortKey="Steinkellner, Herta" sort="Steinkellner, Herta" uniqKey="Steinkellner H" first="Herta" last="Steinkellner">Herta Steinkellner</name></author><author><name sortKey="Scheiner, Otto" sort="Scheiner, Otto" uniqKey="Scheiner O" first="Otto" last="Scheiner">Otto Scheiner</name></author><author><name sortKey="Breiteneder, Heimo" sort="Breiteneder, Heimo" uniqKey="Breiteneder H" first="Heimo" last="Breiteneder">Heimo Breiteneder</name></author></analytic><series><title level="j">Journal of molecular biology</title><idno type="ISSN">0022-2836</idno><imprint><date when="2003" type="published">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Allergens (chemistry)</term><term>Allergens (genetics)</term><term>Allergens (pharmacology)</term><term>Antifungal Agents (chemistry)</term><term>Antifungal Agents (pharmacology)</term><term>Cloning, Molecular (MeSH)</term><term>DNA, Complementary (MeSH)</term><term>Food Hypersensitivity (blood)</term><term>Food Hypersensitivity (etiology)</term><term>Food Hypersensitivity (immunology)</term><term>Fusarium (drug effects)</term><term>Gene Expression Regulation, Plant (MeSH)</term><term>Humans (MeSH)</term><term>Immunoblotting (MeSH)</term><term>Immunoglobulin E (metabolism)</term><term>Malus (MeSH)</term><term>Models, Molecular (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Penicillium (drug effects)</term><term>Peptide Fragments (metabolism)</term><term>Plant Proteins (chemistry)</term><term>Plant Proteins (genetics)</term><term>Plant Proteins (pharmacology)</term><term>Recombinant Proteins (MeSH)</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (MeSH)</term><term>Sweetening Agents (chemistry)</term><term>Sweetening Agents (pharmacology)</term><term>Tobacco (chemistry)</term><term>Tobacco Mosaic Virus (genetics)</term><term>Transfection (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Allergens</term><term>Antifungal Agents</term><term>Plant Proteins</term><term>Sweetening Agents</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Allergens</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Immunoglobulin E</term><term>Peptide Fragments</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Allergens</term><term>Antifungal Agents</term><term>Plant Proteins</term><term>Sweetening Agents</term></keywords><keywords scheme="MESH" qualifier="blood" xml:lang="en"><term>Food Hypersensitivity</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Fusarium</term><term>Penicillium</term></keywords><keywords scheme="MESH" qualifier="etiology" xml:lang="en"><term>Food Hypersensitivity</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Tobacco Mosaic Virus</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Food Hypersensitivity</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>DNA, Complementary</term><term>Gene Expression Regulation, Plant</term><term>Humans</term><term>Immunoblotting</term><term>Malus</term><term>Models, Molecular</term><term>Molecular Sequence Data</term><term>Recombinant Proteins</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Mal d 2 is a thaumatin-like protein and important allergen of apple fruits that is associated with IgE-mediated symptoms in apple allergic individuals. We obtained a full-length cDNA clone of Mal d 2 from RNA isolated from ripe apple (Malus domestica cv. Golden Delicious). The cDNA's open reading frame encodes a protein of 246 amino acid residues including a signal peptide of 24 residues and two putative glycosylation sites. The deduced amino acid sequence of the mature Mal d 2 protein results in a predicted molecular mass of 23,210.9Da and a calculated pI of 4.55. Sequence comparisons and molecular modeling place Mal d 2 among those pathogenesis-related thaumatin-like proteins that contain a conserved acidic cleft. In order to ensure the correct formation of the protein's eight conserved disulfide bridges we expressed Mal d 2 in Nicotiana benthamiana plants by the use of a tobacco mosaic viral vector. Transfected N.benthamiana plants accumulated Mal d 2 to levels of at least 2% of total soluble protein. MALDI-TOF mass spectrometric analyses of the recombinant Mal d 2 and its proteolytic fragments showed that the apple-specific leader peptide was correctly cleaved off by the host plant and that the mature recombinant protein was intact and not glycosylated. Purified recombinant Mal d 2 displayed the ability to bind IgE from apple-allergic individuals equivalent to natural Mal d 2. In addition, the recombinant thaumatin-like Mal d 2 exhibited antifungal activity against Fusarium oxysporum and Penicillium expansum, implying a function in plant defense against fungal pathogens.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12787673</PMID><DateCompleted><Year>2003</Year><Month>07</Month><Day>24</Day></DateCompleted><DateRevised><Year>2019</Year><Month>07</Month><Day>10</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0022-2836</ISSN><JournalIssue CitedMedium="Print"><Volume>329</Volume><Issue>4</Issue><PubDate><Year>2003</Year><Month>Jun</Month><Day>13</Day></PubDate></JournalIssue><Title>Journal of molecular biology</Title><ISOAbbreviation>J Mol Biol</ISOAbbreviation></Journal><ArticleTitle>Plant-based heterologous expression of Mal d 2, a thaumatin-like protein and allergen of apple (Malus domestica), and its characterization as an antifungal protein.</ArticleTitle><Pagination><MedlinePgn>721-30</MedlinePgn></Pagination><Abstract><AbstractText>Mal d 2 is a thaumatin-like protein and important allergen of apple fruits that is associated with IgE-mediated symptoms in apple allergic individuals. We obtained a full-length cDNA clone of Mal d 2 from RNA isolated from ripe apple (Malus domestica cv. Golden Delicious). The cDNA's open reading frame encodes a protein of 246 amino acid residues including a signal peptide of 24 residues and two putative glycosylation sites. The deduced amino acid sequence of the mature Mal d 2 protein results in a predicted molecular mass of 23,210.9Da and a calculated pI of 4.55. Sequence comparisons and molecular modeling place Mal d 2 among those pathogenesis-related thaumatin-like proteins that contain a conserved acidic cleft. In order to ensure the correct formation of the protein's eight conserved disulfide bridges we expressed Mal d 2 in Nicotiana benthamiana plants by the use of a tobacco mosaic viral vector. Transfected N.benthamiana plants accumulated Mal d 2 to levels of at least 2% of total soluble protein. MALDI-TOF mass spectrometric analyses of the recombinant Mal d 2 and its proteolytic fragments showed that the apple-specific leader peptide was correctly cleaved off by the host plant and that the mature recombinant protein was intact and not glycosylated. Purified recombinant Mal d 2 displayed the ability to bind IgE from apple-allergic individuals equivalent to natural Mal d 2. In addition, the recombinant thaumatin-like Mal d 2 exhibited antifungal activity against Fusarium oxysporum and Penicillium expansum, implying a function in plant defense against fungal pathogens.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Krebitz</LastName><ForeName>Monika</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Pathophysiology, University of Vienna, AKH-EBO-3Q, Waehringer Guertel 18-20, 1090, Vienna, Austria.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wagner</LastName><ForeName>Birgit</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>Ferreira</LastName><ForeName>Fatima</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Peterbauer</LastName><ForeName>Clemens</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Campillo</LastName><ForeName>Nuria</ForeName><Initials>N</Initials></Author><Author ValidYN="Y"><LastName>Witty</LastName><ForeName>Michael</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Kolarich</LastName><ForeName>Daniel</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Steinkellner</LastName><ForeName>Herta</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Scheiner</LastName><ForeName>Otto</ForeName><Initials>O</Initials></Author><Author ValidYN="Y"><LastName>Breiteneder</LastName><ForeName>Heimo</ForeName><Initials>H</Initials></Author></AuthorList><Language>eng</Language><DataBankList CompleteYN="Y"><DataBank><DataBankName>GENBANK</DataBankName><AccessionNumberList><AccessionNumber>AJ243427</AccessionNumber></AccessionNumberList></DataBank></DataBankList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>J Mol Biol</MedlineTA><NlmUniqueID>2985088R</NlmUniqueID><ISSNLinking>0022-2836</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000485">Allergens</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000935">Antifungal Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018076">DNA, Complementary</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010446">Peptide Fragments</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant 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MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018076" MajorTopicYN="N">DNA, Complementary</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005512" MajorTopicYN="N">Food Hypersensitivity</DescriptorName><QualifierName UI="Q000097" MajorTopicYN="N">blood</QualifierName><QualifierName UI="Q000209" MajorTopicYN="N">etiology</QualifierName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005670" MajorTopicYN="N">Fusarium</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="N">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" 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IdType="doi">10.1016/s0022-2836(03)00403-0</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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