Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition.
Identifieur interne : 002517 ( Main/Exploration ); précédent : 002516; suivant : 002518Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition.
Auteurs : Debora N. Okamoto [Brésil] ; Lilian C G. Oliveira ; Marcia Y. Kondo ; Maria H S. Cezari ; Zoltán Szeltner ; Tünde Juhász ; Maria A. Juliano ; Lászl Polgár ; Luiz Juliano ; Iuri E. GouveaSource :
- Biological chemistry [ 1437-4315 ] ; 2010.
Descripteurs français
- KwdFr :
- MESH :
- enzymologie : Virus du SRAS.
- métabolisme : Cysteine endopeptidases, Protéines virales, Virus du SRAS.
- Cinétique, Cysteine endopeptidases, Hydrolyse, Protéines virales, Réplication virale.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Cysteine Endopeptidases, Viral Proteins.
- chemical , metabolism : Cysteine Endopeptidases, Viral Proteins.
- enzymology : SARS Virus.
- metabolism : SARS Virus.
- Hydrolysis, Kinetics, Virus Replication.
Abstract
The 3C-like peptidase of the severe acute respiratory syndrome virus (SARS-CoV) is strictly required for viral replication, thus being a potential target for the development of antiviral agents. In contrast to monomeric picornavirus 3C peptidases, SARS-CoV 3CLpro exists in equilibrium between the monomer and dimer forms in solution, and only the dimer is proteolytically active in dilute buffer solutions. In this study, the increase of SARS-CoV 3CLpro peptidase activity in presence of kosmotropic salts and crowding agents is described. The activation followed the Hofmeister series of anions, with two orders of magnitude enhancement in the presence of Na₂SO₄, whereas the crowding agents polyethylene glycol and bovine serum albumin increased the hydrolytic rate up to 3 times. Kinetic determinations of the monomer dimer dissociation constant (K(d)) indicated that activation was a result of a more active dimer, without significant changes in K(d) values. The activation was found to be independent of substrate length and was derived from both k(cat) increase and K(m) decrease. The viral peptidase activation described here could be related to the crowded intracellular environment and indicates a further fine-tuning mechanism for biological control, particularly in the microenvironment of the vesicles that are induced in host cells during positive strand RNA virus infection.
DOI: 10.1515/BC.2010.145
PubMed: 21087086
Affiliations:
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Le document en format XML
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Kondo</name></author><author><name sortKey="Cezari, Maria H S" sort="Cezari, Maria H S" uniqKey="Cezari M" first="Maria H S" last="Cezari">Maria H S. Cezari</name></author><author><name sortKey="Szeltner, Zoltan" sort="Szeltner, Zoltan" uniqKey="Szeltner Z" first="Zoltán" last="Szeltner">Zoltán Szeltner</name></author><author><name sortKey="Juhasz, Tunde" sort="Juhasz, Tunde" uniqKey="Juhasz T" first="Tünde" last="Juhász">Tünde Juhász</name></author><author><name sortKey="Juliano, Maria A" sort="Juliano, Maria A" uniqKey="Juliano M" first="Maria A" last="Juliano">Maria A. Juliano</name></author><author><name sortKey="Polgar, Laszl" sort="Polgar, Laszl" uniqKey="Polgar L" first="Lászl" last="Polgár">Lászl Polgár</name></author><author><name sortKey="Juliano, Luiz" sort="Juliano, Luiz" uniqKey="Juliano L" first="Luiz" last="Juliano">Luiz Juliano</name></author><author><name sortKey="Gouvea, Iuri E" sort="Gouvea, Iuri E" uniqKey="Gouvea I" first="Iuri E" last="Gouvea">Iuri E. 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Okamoto</name><affiliation wicri:level="1"><nlm:affiliation>Departamento de Biofísica, Universidade Federal de São Paulo, Brazil.</nlm:affiliation><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Biofísica, Universidade Federal de São Paulo</wicri:regionArea><wicri:noRegion>Universidade Federal de São Paulo</wicri:noRegion></affiliation></author><author><name sortKey="Oliveira, Lilian C G" sort="Oliveira, Lilian C G" uniqKey="Oliveira L" first="Lilian C G" last="Oliveira">Lilian C G. Oliveira</name></author><author><name sortKey="Kondo, Marcia Y" sort="Kondo, Marcia Y" uniqKey="Kondo M" first="Marcia Y" last="Kondo">Marcia Y. Kondo</name></author><author><name sortKey="Cezari, Maria H S" sort="Cezari, Maria H S" uniqKey="Cezari M" first="Maria H S" last="Cezari">Maria H S. Cezari</name></author><author><name sortKey="Szeltner, Zoltan" sort="Szeltner, Zoltan" uniqKey="Szeltner Z" first="Zoltán" last="Szeltner">Zoltán Szeltner</name></author><author><name sortKey="Juhasz, Tunde" sort="Juhasz, Tunde" uniqKey="Juhasz T" first="Tünde" last="Juhász">Tünde Juhász</name></author><author><name sortKey="Juliano, Maria A" sort="Juliano, Maria A" uniqKey="Juliano M" first="Maria A" last="Juliano">Maria A. Juliano</name></author><author><name sortKey="Polgar, Laszl" sort="Polgar, Laszl" uniqKey="Polgar L" first="Lászl" last="Polgár">Lászl Polgár</name></author><author><name sortKey="Juliano, Luiz" sort="Juliano, Luiz" uniqKey="Juliano L" first="Luiz" last="Juliano">Luiz Juliano</name></author><author><name sortKey="Gouvea, Iuri E" sort="Gouvea, Iuri E" uniqKey="Gouvea I" first="Iuri E" last="Gouvea">Iuri E. Gouvea</name></author></analytic><series><title level="j">Biological chemistry</title><idno type="eISSN">1437-4315</idno><imprint><date when="2010" type="published">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cysteine Endopeptidases (chemistry)</term><term>Cysteine Endopeptidases (metabolism)</term><term>Hydrolysis</term><term>Kinetics</term><term>SARS Virus (enzymology)</term><term>SARS Virus (metabolism)</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (metabolism)</term><term>Virus Replication</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Cinétique</term><term>Cysteine endopeptidases ()</term><term>Cysteine endopeptidases (métabolisme)</term><term>Hydrolyse</term><term>Protéines virales ()</term><term>Protéines virales (métabolisme)</term><term>Réplication virale</term><term>Virus du SRAS (enzymologie)</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Cysteine Endopeptidases</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cysteine Endopeptidases</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cysteine endopeptidases</term><term>Protéines virales</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Hydrolysis</term><term>Kinetics</term><term>Virus Replication</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cinétique</term><term>Cysteine endopeptidases</term><term>Hydrolyse</term><term>Protéines virales</term><term>Réplication virale</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The 3C-like peptidase of the severe acute respiratory syndrome virus (SARS-CoV) is strictly required for viral replication, thus being a potential target for the development of antiviral agents. In contrast to monomeric picornavirus 3C peptidases, SARS-CoV 3CLpro exists in equilibrium between the monomer and dimer forms in solution, and only the dimer is proteolytically active in dilute buffer solutions. In this study, the increase of SARS-CoV 3CLpro peptidase activity in presence of kosmotropic salts and crowding agents is described. The activation followed the Hofmeister series of anions, with two orders of magnitude enhancement in the presence of Na₂SO₄, whereas the crowding agents polyethylene glycol and bovine serum albumin increased the hydrolytic rate up to 3 times. Kinetic determinations of the monomer dimer dissociation constant (K(d)) indicated that activation was a result of a more active dimer, without significant changes in K(d) values. The activation was found to be independent of substrate length and was derived from both k(cat) increase and K(m) decrease. The viral peptidase activation described here could be related to the crowded intracellular environment and indicates a further fine-tuning mechanism for biological control, particularly in the microenvironment of the vesicles that are induced in host cells during positive strand RNA virus infection.</div></front></TEI><affiliations><list><country><li>Brésil</li></country></list><tree><noCountry><name sortKey="Cezari, Maria H S" sort="Cezari, Maria H S" uniqKey="Cezari M" first="Maria H S" last="Cezari">Maria H S. Cezari</name><name sortKey="Gouvea, Iuri E" sort="Gouvea, Iuri E" uniqKey="Gouvea I" first="Iuri E" last="Gouvea">Iuri E. Gouvea</name><name sortKey="Juhasz, Tunde" sort="Juhasz, Tunde" uniqKey="Juhasz T" first="Tünde" last="Juhász">Tünde Juhász</name><name sortKey="Juliano, Luiz" sort="Juliano, Luiz" uniqKey="Juliano L" first="Luiz" last="Juliano">Luiz Juliano</name><name sortKey="Juliano, Maria A" sort="Juliano, Maria A" uniqKey="Juliano M" first="Maria A" last="Juliano">Maria A. Juliano</name><name sortKey="Kondo, Marcia Y" sort="Kondo, Marcia Y" uniqKey="Kondo M" first="Marcia Y" last="Kondo">Marcia Y. Kondo</name><name sortKey="Oliveira, Lilian C G" sort="Oliveira, Lilian C G" uniqKey="Oliveira L" first="Lilian C G" last="Oliveira">Lilian C G. Oliveira</name><name sortKey="Polgar, Laszl" sort="Polgar, Laszl" uniqKey="Polgar L" first="Lászl" last="Polgár">Lászl Polgár</name><name sortKey="Szeltner, Zoltan" sort="Szeltner, Zoltan" uniqKey="Szeltner Z" first="Zoltán" last="Szeltner">Zoltán Szeltner</name></noCountry><country name="Brésil"><noRegion><name sortKey="Okamoto, Debora N" sort="Okamoto, Debora N" uniqKey="Okamoto D" first="Debora N" last="Okamoto">Debora N. Okamoto</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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