The SARS Coronavirus 3a protein binds calcium in its cytoplasmic domain.
Identifieur interne : 001547 ( Main/Exploration ); précédent : 001546; suivant : 001548The SARS Coronavirus 3a protein binds calcium in its cytoplasmic domain.
Auteurs : Rinki Minakshi [Inde] ; Kartika Padhan [Inde] ; Safikur Rehman [Inde] ; Md Imtaiyaz Hassan [Inde] ; Faizan Ahmad [Inde]Source :
- Virus research [ 1872-7492 ] ; 2014.
Descripteurs français
- KwdFr :
- Calcium (métabolisme), Conformation des protéines, Cytoplasme (virologie), Données de séquences moléculaires, Humains, Protéines virales (), Protéines virales (génétique), Protéines virales (métabolisme), Structure tertiaire des protéines, Syndrome respiratoire aigu sévère (virologie), Séquence d'acides aminés, Virus du SRAS (), Virus du SRAS (génétique), Virus du SRAS (métabolisme).
- MESH :
- génétique : Protéines virales, Virus du SRAS.
- métabolisme : Calcium, Protéines virales, Virus du SRAS.
- virologie : Cytoplasme, Syndrome respiratoire aigu sévère.
- Conformation des protéines, Données de séquences moléculaires, Humains, Protéines virales, Structure tertiaire des protéines, Séquence d'acides aminés, Virus du SRAS.
English descriptors
- KwdEn :
- Amino Acid Sequence, Calcium (metabolism), Cytoplasm (virology), Humans, Molecular Sequence Data, Protein Conformation, Protein Structure, Tertiary, SARS Virus (chemistry), SARS Virus (genetics), SARS Virus (metabolism), Severe Acute Respiratory Syndrome (virology), Viral Proteins (chemistry), Viral Proteins (genetics), Viral Proteins (metabolism).
- MESH :
- chemical , chemistry : Viral Proteins.
- chemical , genetics : Viral Proteins.
- chemical , metabolism : Calcium, Viral Proteins.
- chemistry : SARS Virus.
- genetics : SARS Virus.
- metabolism : SARS Virus.
- virology : Cytoplasm, Severe Acute Respiratory Syndrome.
- Amino Acid Sequence, Humans, Molecular Sequence Data, Protein Conformation, Protein Structure, Tertiary.
Abstract
The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) is a positive stranded RNA virus with ∼30kb genome. Among all open reading frames (orfs) of this virus, the orf3a is the largest, and encodes a protein of 274 amino acids, named as 3a protein. Sequence analysis suggests that the orf3a aligned to one calcium pump present in Plasmodium falciparum and the enzyme glutamine synthetase found in Leptospira interrogans. This sequence similarity was found to be limited only to amino acid residues 209-264 which form the cytoplasmic domain of the orf3a. Furthermore, this region was predicted to be involved in the calcium binding. Owing to this hypothesis, we were driven to establish its calcium binding property in vitro. Here, we expressed and purified the cytoplasmic domain of the 3a protein, called Cyto3a, as a recombinant His-tagged protein in the E. coli. The calcium binding nature was established by performing various staining methods such as ruthenium red and stains-all. (45)Ca overlay method was also done to further support the data. Since the 3a protein forms ion channels, we were interested to see any conformational changes occurring in the Cyot3a upon calcium binding, using fluorescence spectroscopy and circular dichroism. These studies clearly indicate a significant change in the conformation of the Cyto3a protein after binding with calcium. Our results strongly suggest that the cytoplasmic domain of the 3a protein of SARS-CoV binds calcium in vitro, causing a change in protein conformation.
DOI: 10.1016/j.virusres.2014.08.001
PubMed: 25116391
Affiliations:
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Electronic address: rinki.minakshi@hotmail.com.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>International Centre for Genetic Engineering and Biotechnology, New Delhi, India; Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi</wicri:regionArea><wicri:noRegion>New Delhi</wicri:noRegion></affiliation></author><author><name sortKey="Padhan, Kartika" sort="Padhan, Kartika" uniqKey="Padhan K" first="Kartika" last="Padhan">Kartika Padhan</name><affiliation wicri:level="1"><nlm:affiliation>International Centre for Genetic Engineering and Biotechnology, New Delhi, India.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>International Centre for Genetic Engineering and Biotechnology, New Delhi</wicri:regionArea><wicri:noRegion>New Delhi</wicri:noRegion></affiliation></author><author><name sortKey="Rehman, Safikur" sort="Rehman, Safikur" uniqKey="Rehman S" first="Safikur" last="Rehman">Safikur Rehman</name><affiliation wicri:level="1"><nlm:affiliation>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, India.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi</wicri:regionArea><wicri:noRegion>New Delhi</wicri:noRegion></affiliation></author><author><name sortKey="Hassan, Md Imtaiyaz" sort="Hassan, Md Imtaiyaz" uniqKey="Hassan M" first="Md Imtaiyaz" last="Hassan">Md Imtaiyaz Hassan</name><affiliation wicri:level="1"><nlm:affiliation>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, India.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi</wicri:regionArea><wicri:noRegion>New Delhi</wicri:noRegion></affiliation></author><author><name sortKey="Ahmad, Faizan" sort="Ahmad, Faizan" uniqKey="Ahmad F" first="Faizan" last="Ahmad">Faizan Ahmad</name><affiliation wicri:level="1"><nlm:affiliation>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, India.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi</wicri:regionArea><wicri:noRegion>New Delhi</wicri:noRegion></affiliation></author></analytic><series><title level="j">Virus research</title><idno type="eISSN">1872-7492</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Calcium (metabolism)</term><term>Cytoplasm (virology)</term><term>Humans</term><term>Molecular Sequence Data</term><term>Protein Conformation</term><term>Protein Structure, Tertiary</term><term>SARS Virus (chemistry)</term><term>SARS Virus (genetics)</term><term>SARS Virus (metabolism)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Calcium (métabolisme)</term><term>Conformation des protéines</term><term>Cytoplasme (virologie)</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Protéines virales ()</term><term>Protéines virales (génétique)</term><term>Protéines virales (métabolisme)</term><term>Structure tertiaire des protéines</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Séquence d'acides aminés</term><term>Virus du SRAS ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Calcium</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines virales</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Calcium</term><term>Protéines virales</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cytoplasme</term><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Cytoplasm</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Humans</term><term>Molecular Sequence Data</term><term>Protein Conformation</term><term>Protein Structure, Tertiary</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Conformation des protéines</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Protéines virales</term><term>Structure tertiaire des protéines</term><term>Séquence d'acides aminés</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) is a positive stranded RNA virus with ∼30kb genome. Among all open reading frames (orfs) of this virus, the orf3a is the largest, and encodes a protein of 274 amino acids, named as 3a protein. Sequence analysis suggests that the orf3a aligned to one calcium pump present in Plasmodium falciparum and the enzyme glutamine synthetase found in Leptospira interrogans. This sequence similarity was found to be limited only to amino acid residues 209-264 which form the cytoplasmic domain of the orf3a. Furthermore, this region was predicted to be involved in the calcium binding. Owing to this hypothesis, we were driven to establish its calcium binding property in vitro. Here, we expressed and purified the cytoplasmic domain of the 3a protein, called Cyto3a, as a recombinant His-tagged protein in the E. coli. The calcium binding nature was established by performing various staining methods such as ruthenium red and stains-all. (45)Ca overlay method was also done to further support the data. Since the 3a protein forms ion channels, we were interested to see any conformational changes occurring in the Cyot3a upon calcium binding, using fluorescence spectroscopy and circular dichroism. These studies clearly indicate a significant change in the conformation of the Cyto3a protein after binding with calcium. Our results strongly suggest that the cytoplasmic domain of the 3a protein of SARS-CoV binds calcium in vitro, causing a change in protein conformation. </div></front></TEI><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Minakshi, Rinki" sort="Minakshi, Rinki" uniqKey="Minakshi R" first="Rinki" last="Minakshi">Rinki Minakshi</name></noRegion><name sortKey="Ahmad, Faizan" sort="Ahmad, Faizan" uniqKey="Ahmad F" first="Faizan" last="Ahmad">Faizan Ahmad</name><name sortKey="Hassan, Md Imtaiyaz" sort="Hassan, Md Imtaiyaz" uniqKey="Hassan M" first="Md Imtaiyaz" last="Hassan">Md Imtaiyaz Hassan</name><name sortKey="Padhan, Kartika" sort="Padhan, Kartika" uniqKey="Padhan K" first="Kartika" last="Padhan">Kartika Padhan</name><name sortKey="Rehman, Safikur" sort="Rehman, Safikur" uniqKey="Rehman S" first="Safikur" last="Rehman">Safikur Rehman</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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