Nucleotide-type chemical shift assignment of the encapsulated 40 kbp dsDNA in intact bacteriophage T7 by MAS solid-state NMR
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作者:
Abramov, Gili
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Tel Aviv Univ, Sch Chem, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, IsraelTel Aviv Univ, Sch Chem, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, Israel
Abramov, Gili
[1
]
Goldbourt, Amir
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Tel Aviv Univ, Sch Chem, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, IsraelTel Aviv Univ, Sch Chem, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, Israel
Goldbourt, Amir
[1
]
机构:
[1] Tel Aviv Univ, Sch Chem, Raymond & Beverly Sackler Fac Exact Sci, IL-69978 Tel Aviv, Israel
The icosahedral bacteriophage T7 is a 50 MDa double-stranded DNA (dsDNA) virus that infects Escherichia coli. Although there is substantial information on the physical and morphological properties of T7, structural information, based mostly on Raman spectroscopy and cryo-electron microscopy, is limited. Here, we apply the magic-angle spinning (MAS) solid-state NMR (SSNMR) technique to study a uniformly C-13 and N-15 labeled wild-type T7 phage. We describe the details of the large-scale preparation and purification of an isotopically enriched phage sample under fully hydrated conditions, and show a complete C-13 and a near-complete N-15 nucleotide-type specific assignment of the sugar and base moieties in the 40 kbp dsDNA of T7 using two-dimensional C-13-C-13 and N-15-C-13 correlation experiments. The chemical shifts are interpreted as reporters of a B-form conformation of the encapsulated dsDNA. While MAS SSNMR was found to be extremely useful in determining the structures of proteins in native-like environments, its application to nucleic acids has lagged behind, leaving a missing C-13 and N-15 chemical shift database. This work therefore expands the C-13 and N-15 database of real B-form DNA systems, and opens routes to characterize more complex nucleic acid systems by SSNMR.
机构:
Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Cady, Sarah D.
;
Schmidt-Rohr, Klaus
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Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Schmidt-Rohr, Klaus
;
Wang, Jun
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Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Wang, Jun
;
Soto, Cinque S.
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机构:
Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Soto, Cinque S.
;
DeGrado, William F.
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机构:
Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
DeGrado, William F.
;
Hong, Mei
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机构:
Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
机构:
Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Cady, Sarah D.
;
Schmidt-Rohr, Klaus
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机构:
Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Schmidt-Rohr, Klaus
;
Wang, Jun
论文数: 0引用数: 0
h-index: 0
机构:
Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Wang, Jun
;
Soto, Cinque S.
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h-index: 0
机构:
Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
Soto, Cinque S.
;
DeGrado, William F.
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机构:
Univ Penn, Sch Med, Dept Biochem & Biophys, Philadelphia, PA 19104 USA
Univ Penn, Dept Chem, Philadelphia, PA 19104 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA
DeGrado, William F.
;
Hong, Mei
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机构:
Iowa State Univ, Dept Chem, Ames, IA 50011 USAIowa State Univ, Dept Chem, Ames, IA 50011 USA