Probing the sweet determinants of brazzein: wild-type brazzein and a tasteless variant, brazzein-ins(R18a-I18b), exhibit different pH-dependent NMR chemical shifts.
Identifieur interne : 000412 ( Main/Corpus ); précédent : 000411; suivant : 000413Probing the sweet determinants of brazzein: wild-type brazzein and a tasteless variant, brazzein-ins(R18a-I18b), exhibit different pH-dependent NMR chemical shifts.
Auteurs : Qin Zhao ; Jikui Song ; Zheyuan Jin ; Vicktoria Danilova ; Göran Hellekant ; John L. MarkleySource :
- Biochemical and biophysical research communications [ 0006-291X ] ; 2005.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Plant Proteins, Sweetening Agents.
- genetics : Genetic Variation, Plant Proteins.
- Hydrogen-Ion Concentration, Models, Molecular, Nuclear Magnetic Resonance, Biomolecular, Protein Structure, Tertiary.
Abstract
Brazzein is a small, intensely sweet protein. As a probe of the functional properties of its solvent-exposed loop, two residues (Arg-Ile) were inserted between Leu18 and Ala19 of brazzein. Psychophysical testing demonstrated that this mutant is totally tasteless. NMR chemical shift mapping of differences between this mutant and brazzein indicated that residues affected by the insertion are localized to the mutated loop, the region of the single alpha-helix, and around the Cys16-Cys37 disulfide bond. Residues unaffected by this mutation included those near the C-terminus and in the loop connecting the alpha-helix and the second beta-strand. In particular, several residues of brazzein previously shown to be essential for its sweetness (His31, Arg33, Glu41, Arg43, Asp50, and Tyr54) exhibited negligible chemical shift changes. Moreover, the pH dependence of the chemical shifts of His31, Glu41, Asp50, and Tyr54 were unaltered by the insertion. The insertion led to large chemical shift and pKa perturbation of Glu36, a residue shown previously to be important for brazzein's sweetness. These results serve to refine the known sweetness determinants of brazzein and lend further support to the idea that the protein interacts with a sweet-taste receptor through a multi-site interaction mechanism, as has been postulated for brazzein and other sweet proteins (monellin and thaumatin).
DOI: 10.1016/j.bbrc.2005.07.049
PubMed: 16105551
Links to Exploration step
pubmed:16105551Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Probing the sweet determinants of brazzein: wild-type brazzein and a tasteless variant, brazzein-ins(R18a-I18b), exhibit different pH-dependent NMR chemical shifts.</title><author><name sortKey="Zhao, Qin" sort="Zhao, Qin" uniqKey="Zhao Q" first="Qin" last="Zhao">Qin Zhao</name><affiliation><nlm:affiliation>Department of Biochemistry, University of Wisconsin Madison, College of Agricultural and Life Sciences, USA. qin@nmrfam.wisc.edu</nlm:affiliation></affiliation></author><author><name sortKey="Song, Jikui" sort="Song, Jikui" uniqKey="Song J" first="Jikui" last="Song">Jikui Song</name></author><author><name sortKey="Jin, Zheyuan" sort="Jin, Zheyuan" uniqKey="Jin Z" first="Zheyuan" last="Jin">Zheyuan Jin</name></author><author><name sortKey="Danilova, Vicktoria" sort="Danilova, Vicktoria" uniqKey="Danilova V" first="Vicktoria" last="Danilova">Vicktoria Danilova</name></author><author><name sortKey="Hellekant, Goran" sort="Hellekant, Goran" uniqKey="Hellekant G" first="Göran" last="Hellekant">Göran Hellekant</name></author><author><name sortKey="Markley, John L" sort="Markley, John L" uniqKey="Markley J" first="John L" last="Markley">John L. Markley</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16105551</idno><idno type="pmid">16105551</idno><idno type="doi">10.1016/j.bbrc.2005.07.049</idno><idno type="wicri:Area/Main/Corpus">000412</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000412</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Probing the sweet determinants of brazzein: wild-type brazzein and a tasteless variant, brazzein-ins(R18a-I18b), exhibit different pH-dependent NMR chemical shifts.</title><author><name sortKey="Zhao, Qin" sort="Zhao, Qin" uniqKey="Zhao Q" first="Qin" last="Zhao">Qin Zhao</name><affiliation><nlm:affiliation>Department of Biochemistry, University of Wisconsin Madison, College of Agricultural and Life Sciences, USA. qin@nmrfam.wisc.edu</nlm:affiliation></affiliation></author><author><name sortKey="Song, Jikui" sort="Song, Jikui" uniqKey="Song J" first="Jikui" last="Song">Jikui Song</name></author><author><name sortKey="Jin, Zheyuan" sort="Jin, Zheyuan" uniqKey="Jin Z" first="Zheyuan" last="Jin">Zheyuan Jin</name></author><author><name sortKey="Danilova, Vicktoria" sort="Danilova, Vicktoria" uniqKey="Danilova V" first="Vicktoria" last="Danilova">Vicktoria Danilova</name></author><author><name sortKey="Hellekant, Goran" sort="Hellekant, Goran" uniqKey="Hellekant G" first="Göran" last="Hellekant">Göran Hellekant</name></author><author><name sortKey="Markley, John L" sort="Markley, John L" uniqKey="Markley J" first="John L" last="Markley">John L. Markley</name></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="ISSN">0006-291X</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Genetic Variation (genetics)</term><term>Hydrogen-Ion Concentration (MeSH)</term><term>Models, Molecular (MeSH)</term><term>Nuclear Magnetic Resonance, Biomolecular (MeSH)</term><term>Plant Proteins (chemistry)</term><term>Plant Proteins (genetics)</term><term>Protein Structure, Tertiary (MeSH)</term><term>Sweetening Agents (chemistry)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Plant Proteins</term><term>Sweetening Agents</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Genetic Variation</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Hydrogen-Ion Concentration</term><term>Models, Molecular</term><term>Nuclear Magnetic Resonance, Biomolecular</term><term>Protein Structure, Tertiary</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Brazzein is a small, intensely sweet protein. As a probe of the functional properties of its solvent-exposed loop, two residues (Arg-Ile) were inserted between Leu18 and Ala19 of brazzein. Psychophysical testing demonstrated that this mutant is totally tasteless. NMR chemical shift mapping of differences between this mutant and brazzein indicated that residues affected by the insertion are localized to the mutated loop, the region of the single alpha-helix, and around the Cys16-Cys37 disulfide bond. Residues unaffected by this mutation included those near the C-terminus and in the loop connecting the alpha-helix and the second beta-strand. In particular, several residues of brazzein previously shown to be essential for its sweetness (His31, Arg33, Glu41, Arg43, Asp50, and Tyr54) exhibited negligible chemical shift changes. Moreover, the pH dependence of the chemical shifts of His31, Glu41, Asp50, and Tyr54 were unaltered by the insertion. The insertion led to large chemical shift and pKa perturbation of Glu36, a residue shown previously to be important for brazzein's sweetness. These results serve to refine the known sweetness determinants of brazzein and lend further support to the idea that the protein interacts with a sweet-taste receptor through a multi-site interaction mechanism, as has been postulated for brazzein and other sweet proteins (monellin and thaumatin).</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16105551</PMID><DateCompleted><Year>2005</Year><Month>10</Month><Day>11</Day></DateCompleted><DateRevised><Year>2008</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-291X</ISSN><JournalIssue CitedMedium="Print"><Volume>335</Volume><Issue>1</Issue><PubDate><Year>2005</Year><Month>Sep</Month><Day>16</Day></PubDate></JournalIssue><Title>Biochemical and biophysical research communications</Title><ISOAbbreviation>Biochem Biophys Res Commun</ISOAbbreviation></Journal><ArticleTitle>Probing the sweet determinants of brazzein: wild-type brazzein and a tasteless variant, brazzein-ins(R18a-I18b), exhibit different pH-dependent NMR chemical shifts.</ArticleTitle><Pagination><MedlinePgn>256-63</MedlinePgn></Pagination><Abstract><AbstractText>Brazzein is a small, intensely sweet protein. As a probe of the functional properties of its solvent-exposed loop, two residues (Arg-Ile) were inserted between Leu18 and Ala19 of brazzein. Psychophysical testing demonstrated that this mutant is totally tasteless. NMR chemical shift mapping of differences between this mutant and brazzein indicated that residues affected by the insertion are localized to the mutated loop, the region of the single alpha-helix, and around the Cys16-Cys37 disulfide bond. Residues unaffected by this mutation included those near the C-terminus and in the loop connecting the alpha-helix and the second beta-strand. In particular, several residues of brazzein previously shown to be essential for its sweetness (His31, Arg33, Glu41, Arg43, Asp50, and Tyr54) exhibited negligible chemical shift changes. Moreover, the pH dependence of the chemical shifts of His31, Glu41, Asp50, and Tyr54 were unaltered by the insertion. The insertion led to large chemical shift and pKa perturbation of Glu36, a residue shown previously to be important for brazzein's sweetness. These results serve to refine the known sweetness determinants of brazzein and lend further support to the idea that the protein interacts with a sweet-taste receptor through a multi-site interaction mechanism, as has been postulated for brazzein and other sweet proteins (monellin and thaumatin).</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhao</LastName><ForeName>Qin</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, University of Wisconsin Madison, College of Agricultural and Life Sciences, USA. qin@nmrfam.wisc.edu</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Song</LastName><ForeName>Jikui</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Jin</LastName><ForeName>Zheyuan</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Danilova</LastName><ForeName>Vicktoria</ForeName><Initials>V</Initials></Author><Author ValidYN="Y"><LastName>Hellekant</LastName><ForeName>Göran</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Markley</LastName><ForeName>John L</ForeName><Initials>JL</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>P41 RR02301</GrantID><Acronym>RR</Acronym><Agency>NCRR NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>R01 DC006016</GrantID><Acronym>DC</Acronym><Agency>NIDCD NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D052061">Research Support, N.I.H., Extramural</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biochem Biophys Res Commun</MedlineTA><NlmUniqueID>0372516</NlmUniqueID><ISSNLinking>0006-291X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D013549">Sweetening Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C090521">brazzein protein, Pentadiplandra brazzeana</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D014644" MajorTopicYN="N">Genetic Variation</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008958" MajorTopicYN="N">Models, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019906" MajorTopicYN="N">Nuclear Magnetic Resonance, Biomolecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017434" MajorTopicYN="N">Protein Structure, Tertiary</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013549" MajorTopicYN="N">Sweetening Agents</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2005</Year><Month>06</Month><Day>27</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2005</Year><Month>07</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2005</Year><Month>8</Month><Day>18</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>10</Month><Day>12</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>8</Month><Day>18</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">16105551</ArticleId><ArticleId IdType="pii">S0006-291X(05)01450-6</ArticleId><ArticleId IdType="doi">10.1016/j.bbrc.2005.07.049</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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