Abstract
Single-chain monellin (SCM), which is an engineered 94-residue polypeptide, has proven to be as sweet as native two-chain monellin. SCM is more stable than the native monellin for both heat and acidic environments. Data from gel filtration HPLC and NMR indicate that the SCM exists as a monomer in aqueous solution. The solution structure of SCM has been determined by nuclear magnetic resonance (NMR) spectroscopy and dynamical simulated annealing calculations. A stable α-helix spanning residues Phe11- Ile26 and an antiparallel β-sheet formed by residues 2-5, 36-38, 41-47, 54- 64, 69-75, and 83-88 have been identified. The sheet was well defined by backbone-backbone NOEs, and the corresponding β-strands were further confirmed by hydrogen bond networks based on amide hydrogen exchange data. Strands β2 and β3 are connected by a small bulge comprising residues Ile38- Cys41. A total of 993 distance and 56 dihedral angle restraints were used for simulated annealing calculations. The final simulated annealing structures (<SA>(k)) converged well with a root-mean-square deviation (rmsd) between backbone atoms of 0.49 Å for secondary structural regions and 0.70 Å for backbone atoms excluding two loop regions. The average restraint energy- minimized (REM) structure exhibited root-mean-square deviations of 1.19 Å for backbone atoms and 0.85 Å for backbone atoms excluding two loop regions with respect to 20 <SA>(k) structures. The solution structure of SCM revealed that the long α-helix was folded into the concave side of a six-stranded antiparallel β-sheet. The side chains of Tyr63 and Asp66 which are common to all sweet peptides showed an opposite orientation relative to H1 helix, and they were all solvent-exposed. Residues at the proposed dimeric interface in the X-ray structure were observed to be mostly solvent-exposed and demonstrated high degrees of flexibility.
| Original language | English |
|---|---|
| Pages (from-to) | 2340-2346 |
| Number of pages | 7 |
| Journal | Biochemistry |
| Volume | 38 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 1999 Feb 23 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
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Solution structure of a sweet protein single-chain monellin determined by nuclear magnetic resonance and dynamical simulated annealing calculations. / Lee, Seok Yong; Lee, Jung Hoon; Chang, Ho Jin; Cho, Joong Myung; Jung, Jin Won; Lee, Weon Tae.
In: Biochemistry, Vol. 38, No. 8, 23.02.1999, p. 2340-2346.Research output: Contribution to journal › Article
TY - JOUR
T1 - Solution structure of a sweet protein single-chain monellin determined by nuclear magnetic resonance and dynamical simulated annealing calculations
AU - Lee, Seok Yong
AU - Lee, Jung Hoon
AU - Chang, Ho Jin
AU - Cho, Joong Myung
AU - Jung, Jin Won
AU - Lee, Weon Tae
PY - 1999/2/23
Y1 - 1999/2/23
N2 - Single-chain monellin (SCM), which is an engineered 94-residue polypeptide, has proven to be as sweet as native two-chain monellin. SCM is more stable than the native monellin for both heat and acidic environments. Data from gel filtration HPLC and NMR indicate that the SCM exists as a monomer in aqueous solution. The solution structure of SCM has been determined by nuclear magnetic resonance (NMR) spectroscopy and dynamical simulated annealing calculations. A stable α-helix spanning residues Phe11- Ile26 and an antiparallel β-sheet formed by residues 2-5, 36-38, 41-47, 54- 64, 69-75, and 83-88 have been identified. The sheet was well defined by backbone-backbone NOEs, and the corresponding β-strands were further confirmed by hydrogen bond networks based on amide hydrogen exchange data. Strands β2 and β3 are connected by a small bulge comprising residues Ile38- Cys41. A total of 993 distance and 56 dihedral angle restraints were used for simulated annealing calculations. The final simulated annealing structures (<SA>(k)) converged well with a root-mean-square deviation (rmsd) between backbone atoms of 0.49 Å for secondary structural regions and 0.70 Å for backbone atoms excluding two loop regions. The average restraint energy- minimized (REM) structure exhibited root-mean-square deviations of 1.19 Å for backbone atoms and 0.85 Å for backbone atoms excluding two loop regions with respect to 20 <SA>(k) structures. The solution structure of SCM revealed that the long α-helix was folded into the concave side of a six-stranded antiparallel β-sheet. The side chains of Tyr63 and Asp66 which are common to all sweet peptides showed an opposite orientation relative to H1 helix, and they were all solvent-exposed. Residues at the proposed dimeric interface in the X-ray structure were observed to be mostly solvent-exposed and demonstrated high degrees of flexibility.
AB - Single-chain monellin (SCM), which is an engineered 94-residue polypeptide, has proven to be as sweet as native two-chain monellin. SCM is more stable than the native monellin for both heat and acidic environments. Data from gel filtration HPLC and NMR indicate that the SCM exists as a monomer in aqueous solution. The solution structure of SCM has been determined by nuclear magnetic resonance (NMR) spectroscopy and dynamical simulated annealing calculations. A stable α-helix spanning residues Phe11- Ile26 and an antiparallel β-sheet formed by residues 2-5, 36-38, 41-47, 54- 64, 69-75, and 83-88 have been identified. The sheet was well defined by backbone-backbone NOEs, and the corresponding β-strands were further confirmed by hydrogen bond networks based on amide hydrogen exchange data. Strands β2 and β3 are connected by a small bulge comprising residues Ile38- Cys41. A total of 993 distance and 56 dihedral angle restraints were used for simulated annealing calculations. The final simulated annealing structures (<SA>(k)) converged well with a root-mean-square deviation (rmsd) between backbone atoms of 0.49 Å for secondary structural regions and 0.70 Å for backbone atoms excluding two loop regions. The average restraint energy- minimized (REM) structure exhibited root-mean-square deviations of 1.19 Å for backbone atoms and 0.85 Å for backbone atoms excluding two loop regions with respect to 20 <SA>(k) structures. The solution structure of SCM revealed that the long α-helix was folded into the concave side of a six-stranded antiparallel β-sheet. The side chains of Tyr63 and Asp66 which are common to all sweet peptides showed an opposite orientation relative to H1 helix, and they were all solvent-exposed. Residues at the proposed dimeric interface in the X-ray structure were observed to be mostly solvent-exposed and demonstrated high degrees of flexibility.
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U2 - 10.1021/bi9822731
DO - 10.1021/bi9822731
M3 - Article
C2 - 10029527
AN - SCOPUS:0033596696
VL - 38
SP - 2340
EP - 2346
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 8
ER -