Related ArticlesStructural and dynamic properties of the Fv fragment and the single-chain Fv fragment of an antibody in solution investigated by heteronuclear three-dimensional NMR spectroscopy.
Biochemistry. 1994 Mar 22;33(11):3296-303
Authors: Freund C, Ross A, Plückthun A, Holak TA
Fv fragments, heterodimers of the variable light (VL) and variable heavy chain (VH) domains, are the smallest functional antibody units with molecular masses of approximately 26 kDa. The structural and dynamic properties of the Fv fragment and the corresponding single-chain Fv fragment (scFv: VH-linker-VL, 252 amino acids) of the phosphorylcholine-binding antibody McPC603 in the presence of hapten have been studied in solution by heteronuclear multidimensional NMR spectroscopy. Both 15N TOCSY-HMQC and triple-resonance experiments (HNCA and HN(CA)H, with 15N-13C-labeled protein) gave poor spectra, due to short T2 relaxation times for most of the backbone 1H, 15N, and 13C alpha atoms. The assignment procedure therefore relied upon the combination of amino acid and domain (VL) specifically labeled spectra and the 3D NOESY-HMQC spectrum of the uniformly 15N labeled Fv and scFv fragments. Approximately 80% of the 15N and 1H backbone and 60% of the 1H side-chain resonances have been assigned. Short- and long-range NOEs were used to determine the extent of beta-sheet structure and were compared to the X-ray crystallographic data. The 1H-15N NOE data indicate that the scFv backbone has a well-defined structure of limited conformational flexibility. However, the linker of the scFv fragment exhibits substantial fast internal motion (on the picosecond to nanosecond time scale) compared with the overall rotational correlation time of the whole molecule. Several residues in the CDRs, in turns, or at the C-terminal end of the protein have smaller NOEs, reflecting some degree of rapid motion in the protein backbone.
Target immobilization as a strategy for NMR-based fragment screening: comparison of TINS, STD, and SPR for fragment hit identification.
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J Biomol Screen. 2010 Sep;15(8):978-89
Authors: Kobayashi M, Retra K, Figaroa F, Hollander JG, Ab E, Heetebrij RJ, Irth H, Siegal G
Fragment-based drug discovery (FBDD) has become a widely accepted tool that is complementary to high-throughput screening (HTS) in developing...
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[NMR paper] Interactions of a didomain fragment of the Drosophila sex-lethal protein with single-
Interactions of a didomain fragment of the Drosophila sex-lethal protein with single-stranded uridine-rich oligoribonucleotides derived from the transformer and Sex-lethal messenger RNA precursors: NMR with residue-selective uridine substitutions.
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J Biomol NMR. 2000 Jun;17(2):153-65
Authors: Kim...
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[NMR paper] Structure and dynamic properties of the single disulfide-deficient alpha-amylase inhi
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Proteins. 1998 Nov 1;33(2):285-94
Authors: Balbach J, Seip S, Kessler H, Scharf M, Kashani-Poor N, Engels JW
Covalent linkages such as disulfide bonds are important for the stabilization of proteins. In the present NMR study we compare the structure and the dynamics of the single...
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[NMR paper] Helix formation by the phospholipase A2 38-59 fragment: influence of chain shortening
Helix formation by the phospholipase A2 38-59 fragment: influence of chain shortening and dimerization monitored by nmr chemical shifts.
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Biopolymers. 1994 May;34(5):647-61
Authors: Jiménez MA, Carreño C, Andreu D, Blanco FJ, Herranz J, Rico M, Nieto JL
The solution structure of a peptide fragment corresponding to the 38-59 region of porcine phospholipase A2 has been investigated using CD,...
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[NMR paper] Structural analysis of peptide fragment 71-93 of transthyretin by NMR spectroscopy an
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Biochemistry. 1994 Jan 11;33(1):33-41
Authors: Jarvis JA, Munro SL, Craik DJ
A peptide corresponding to the amino acid region 71-93 of the plasma protein transthyretin (TTR) has been synthesized to investigate its role in the native...
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[NMR paper] Helix formation by the phospholipase A2 38-59 fragment: influence of chain shortening
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Biopolymers. 1994 May;34(5):647-61
Authors: Jiménez MA, Carreño C, Andreu D, Blanco FJ, Herranz J, Rico M, Nieto JL
The solution structure of a peptide fragment corresponding to the 38-59 region of porcine phospholipase A2 has been investigated using CD,...
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08-22-2010 03:33 AM
[NMR paper] Structural and dynamic properties of the Fv fragment and the single-chain Fv fragment
Structural and dynamic properties of the Fv fragment and the single-chain Fv fragment of an antibody in solution investigated by heteronuclear three-dimensional NMR spectroscopy.
Related Articles Structural and dynamic properties of the Fv fragment and the single-chain Fv fragment of an antibody in solution investigated by heteronuclear three-dimensional NMR spectroscopy.
Biochemistry. 1994 Mar 22;33(11):3296-303
Authors: Freund C, Ross A, Plückthun A, Holak TA
Fv fragments, heterodimers of the variable light (VL) and variable heavy chain...
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Journal club
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08-22-2010 03:33 AM
[NMR paper] Structural analysis of peptide fragment 71-93 of transthyretin by NMR spectroscopy an
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Biochemistry. 1994 Jan 11;33(1):33-41
Authors: Jarvis JA, Munro SL, Craik DJ
A peptide corresponding to the amino acid region 71-93 of the plasma protein transthyretin (TTR) has been synthesized to investigate its role in the native...