A fluorogenic peptide containing the processing site of human SARS corona virus S-protein: kinetic evaluation and NMR structure elucidation.
Identifieur interne : 000095 ( Ncbi/Checkpoint ); précédent : 000094; suivant : 000096A fluorogenic peptide containing the processing site of human SARS corona virus S-protein: kinetic evaluation and NMR structure elucidation.
Auteurs : Ajoy Basak [Canada] ; Abhijit Mitra ; Sarmistha Basak ; Carolyn Pasko ; Michel Chrétien ; Pamela SeatonSource :
- Chembiochem : a European journal of chemical biology [ 1439-4227 ] ; 2007.
Descripteurs français
- KwdFr :
- Acides aminés (analyse), Algorithmes, Chromatographie en phase liquide à haute performance, Cinétique, Colorants fluorescents, Dichroïsme circulaire, Furanes (), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Peptides (), Protéines de l'enveloppe virale (), Relation quantitative structure-activité, Spectrométrie de masse, Spectroscopie par résonance magnétique, Virus du SRAS ().
- MESH :
- analyse : Acides aminés.
- Algorithmes, Chromatographie en phase liquide à haute performance, Cinétique, Colorants fluorescents, Dichroïsme circulaire, Furanes, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Peptides, Protéines de l'enveloppe virale, Relation quantitative structure-activité, Spectrométrie de masse, Spectroscopie par résonance magnétique, Virus du SRAS.
English descriptors
- KwdEn :
- Algorithms, Amino Acids (analysis), Chromatography, High Pressure Liquid, Circular Dichroism, Fluorescent Dyes, Furans (chemistry), Kinetics, Magnetic Resonance Spectroscopy, Mass Spectrometry, Membrane Glycoproteins (chemistry), Peptides (chemistry), Quantitative Structure-Activity Relationship, SARS Virus (chemistry), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry).
- MESH :
- chemical , analysis : Amino Acids.
- chemical , chemistry : Furans, Membrane Glycoproteins, Peptides, Viral Envelope Proteins.
- chemistry : SARS Virus.
- Algorithms, Chromatography, High Pressure Liquid, Circular Dichroism, Fluorescent Dyes, Kinetics, Magnetic Resonance Spectroscopy, Mass Spectrometry, Quantitative Structure-Activity Relationship, Spike Glycoprotein, Coronavirus.
Abstract
Human severe acute respiratory syndrome coronavirus (hSARS-CoV) is the causative agent for SARS infection. Its surface glycoprotein (spike protein) is considered to be one of the prime targets for SARS therapeutics and intervention because its proteolytic maturation by a host protease is crucial for host-virus fusion. Using intramolecularly quenched fluorogenic (IQF) peptides based on hSARS-CoV spike protein (Abz-(755)Glu-Gln-Asp-Arg-Asn-Thr-Arg-Glu-Val-Phe-Ala-Gln(766)-Tyx-NH(2)) and in vitro studies, we show that besides furin, other PCs, like PC5 and PC7, might also be involved in this cleavage event. Through kinetic measurements with recombinant PCs, we observed that the peptide was cleaved efficiently by both furin and PC5, but very poorly by PC7. The cleavage could be blocked by a PC-inhibitor, alpha1-PDX, in a dose-dependent manner. Circular dichroism spectra indicated that this peptide possesses a high degree of sheet structure. Following cleavage by furin, the sheet content increased, possibly at the expense of turn and random structures. (1)H NMR spectra from 2D COSY and ROESY experiments under physiological buffer and pH conditions indicated that this peptide possesses a structure with a turn at its C-terminal segment, close to the cleavage site. The data suggest that the cleavable peptide bond is located within the most exposed domain; this is supported by the nearby turn structure. Several strong to weak NMR ROESY correlations were detected, and a 3D structure of the spike IQF peptide that contains the crucial cleavage site R(761) E has been proposed.
DOI: 10.1002/cbic.200700007
PubMed: 17471479
Affiliations:
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Mitra</name></author><author><name sortKey="Basak, Sarmistha" sort="Basak, Sarmistha" uniqKey="Basak S" first="Sarmistha" last="Basak">Sarmistha Basak</name></author><author><name sortKey="Pasko, Carolyn" sort="Pasko, Carolyn" uniqKey="Pasko C" first="Carolyn" last="Pasko">Carolyn Pasko</name></author><author><name sortKey="Chretien, Michel" sort="Chretien, Michel" uniqKey="Chretien M" first="Michel" last="Chrétien">Michel Chrétien</name></author><author><name sortKey="Seaton, Pamela" sort="Seaton, Pamela" uniqKey="Seaton P" first="Pamela" last="Seaton">Pamela Seaton</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2007">2007</date><idno type="RBID">pubmed:17471479</idno><idno type="pmid">17471479</idno><idno type="doi">10.1002/cbic.200700007</idno><idno type="wicri:Area/PubMed/Corpus">000692</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000692</idno><idno 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Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Hormone, Growth, and Development Program, Regional Protein Chemistry Center, Ottawa Health Research Institute, University of Ottawa, 725 Parkdale Ave., Ottawa, ON K1Y 4E9</wicri:regionArea><wicri:noRegion>ON K1Y 4E9</wicri:noRegion></affiliation></author><author><name sortKey="Mitra, Abhijit" sort="Mitra, Abhijit" uniqKey="Mitra A" first="Abhijit" last="Mitra">Abhijit Mitra</name></author><author><name sortKey="Basak, Sarmistha" sort="Basak, Sarmistha" uniqKey="Basak S" first="Sarmistha" last="Basak">Sarmistha Basak</name></author><author><name sortKey="Pasko, Carolyn" sort="Pasko, Carolyn" uniqKey="Pasko C" first="Carolyn" last="Pasko">Carolyn Pasko</name></author><author><name sortKey="Chretien, Michel" sort="Chretien, Michel" uniqKey="Chretien M" first="Michel" last="Chrétien">Michel Chrétien</name></author><author><name sortKey="Seaton, Pamela" sort="Seaton, Pamela" uniqKey="Seaton P" first="Pamela" last="Seaton">Pamela Seaton</name></author></analytic><series><title level="j">Chembiochem : a European journal of chemical biology</title><idno type="ISSN">1439-4227</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Algorithms</term><term>Amino Acids (analysis)</term><term>Chromatography, High Pressure Liquid</term><term>Circular Dichroism</term><term>Fluorescent Dyes</term><term>Furans (chemistry)</term><term>Kinetics</term><term>Magnetic Resonance Spectroscopy</term><term>Mass Spectrometry</term><term>Membrane Glycoproteins (chemistry)</term><term>Peptides (chemistry)</term><term>Quantitative Structure-Activity Relationship</term><term>SARS Virus (chemistry)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (chemistry)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acides aminés (analyse)</term><term>Algorithmes</term><term>Chromatographie en phase liquide à haute performance</term><term>Cinétique</term><term>Colorants fluorescents</term><term>Dichroïsme circulaire</term><term>Furanes ()</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires ()</term><term>Peptides ()</term><term>Protéines de l'enveloppe virale ()</term><term>Relation quantitative structure-activité</term><term>Spectrométrie de masse</term><term>Spectroscopie par résonance magnétique</term><term>Virus du SRAS ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Amino Acids</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Furans</term><term>Membrane Glycoproteins</term><term>Peptides</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Acides aminés</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Algorithms</term><term>Chromatography, High Pressure Liquid</term><term>Circular Dichroism</term><term>Fluorescent Dyes</term><term>Kinetics</term><term>Magnetic Resonance Spectroscopy</term><term>Mass Spectrometry</term><term>Quantitative Structure-Activity Relationship</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Algorithmes</term><term>Chromatographie en phase liquide à haute performance</term><term>Cinétique</term><term>Colorants fluorescents</term><term>Dichroïsme circulaire</term><term>Furanes</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Peptides</term><term>Protéines de l'enveloppe virale</term><term>Relation quantitative structure-activité</term><term>Spectrométrie de masse</term><term>Spectroscopie par résonance magnétique</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human severe acute respiratory syndrome coronavirus (hSARS-CoV) is the causative agent for SARS infection. Its surface glycoprotein (spike protein) is considered to be one of the prime targets for SARS therapeutics and intervention because its proteolytic maturation by a host protease is crucial for host-virus fusion. Using intramolecularly quenched fluorogenic (IQF) peptides based on hSARS-CoV spike protein (Abz-(755)Glu-Gln-Asp-Arg-Asn-Thr-Arg-Glu-Val-Phe-Ala-Gln(766)-Tyx-NH(2)) and in vitro studies, we show that besides furin, other PCs, like PC5 and PC7, might also be involved in this cleavage event. Through kinetic measurements with recombinant PCs, we observed that the peptide was cleaved efficiently by both furin and PC5, but very poorly by PC7. The cleavage could be blocked by a PC-inhibitor, alpha1-PDX, in a dose-dependent manner. Circular dichroism spectra indicated that this peptide possesses a high degree of sheet structure. Following cleavage by furin, the sheet content increased, possibly at the expense of turn and random structures. (1)H NMR spectra from 2D COSY and ROESY experiments under physiological buffer and pH conditions indicated that this peptide possesses a structure with a turn at its C-terminal segment, close to the cleavage site. The data suggest that the cleavable peptide bond is located within the most exposed domain; this is supported by the nearby turn structure. Several strong to weak NMR ROESY correlations were detected, and a 3D structure of the spike IQF peptide that contains the crucial cleavage site R(761) E has been proposed.</div></front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Basak, Sarmistha" sort="Basak, Sarmistha" uniqKey="Basak S" first="Sarmistha" last="Basak">Sarmistha Basak</name><name sortKey="Chretien, Michel" sort="Chretien, Michel" uniqKey="Chretien M" first="Michel" last="Chrétien">Michel Chrétien</name><name sortKey="Mitra, Abhijit" sort="Mitra, Abhijit" uniqKey="Mitra A" first="Abhijit" last="Mitra">Abhijit Mitra</name><name sortKey="Pasko, Carolyn" sort="Pasko, Carolyn" uniqKey="Pasko C" first="Carolyn" last="Pasko">Carolyn Pasko</name><name sortKey="Seaton, Pamela" sort="Seaton, Pamela" uniqKey="Seaton P" first="Pamela" last="Seaton">Pamela Seaton</name></noCountry><country name="Canada"><noRegion><name sortKey="Basak, Ajoy" sort="Basak, Ajoy" uniqKey="Basak A" first="Ajoy" last="Basak">Ajoy Basak</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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