抄録
A number of studies have examined the structural properties of late folding intermediates of (β/α)8-barrel proteins involved in tryptophan biosynthesis, whereas there is little information available about the early folding events of these proteins. To identify the contiguous polypeptide segments important to the folding of the (β/α)8-barrel protein Escherichia coli N-(5′-phosphoribosyl)anthranilate isomerase, we structurally characterized fragments and circularly permuted forms of the protein. We also simulated thermal unfolding of the protein using molecular dynamics. Our fragmentation experiments demonstrate that the isolated (β/α)1-4β5 fragment is almost as stable as the full-length protein. The far and near-UV CD spectra of this fragment are indicative of native-like secondary and tertiary structures. Structural analysis of the circularly permutated proteins shows that if the protein is cleaved within the two N-terminal βα modules, the amount of secondary structure is unaffected, whereas, when cleaved within the central (β/α)3-4β5 segment, the protein simply cannot fold. An ensemble of the denatured structures produced by thermal unfolding simulations contains a persistent local structure comprised of β3, β4 and β5. The presence of this three-stranded β-barrel suggests that it may be an important early-stage folding intermediate. Interactions found in (β/α) 3-4β5 may be essential for the early events of ePRAI folding if they provide a nucleation site that directs folding.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 1161-1170 |
| ページ数 | 10 |
| ジャーナル | Journal of Molecular Biology |
| 巻 | 353 |
| 発行部数 | 5 |
| DOI | |
| 出版物ステータス | Published - 2005 11 11 |
| 外部発表 | Yes |
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ASJC Scopus subject areas
- Virology
これを引用
Identification and characterization of key substructures involved in the early folding events of a (β/α)8-barrel protein as studied by experimental and computational methods. / Akanuma, Satoshi; Yamagishi, Akihiko.
：: Journal of Molecular Biology, 巻 353, 番号 5, 11.11.2005, p. 1161-1170.研究成果: Article
}
TY - JOUR
T1 - Identification and characterization of key substructures involved in the early folding events of a (β/α)8-barrel protein as studied by experimental and computational methods
AU - Akanuma, Satoshi
AU - Yamagishi, Akihiko
PY - 2005/11/11
Y1 - 2005/11/11
N2 - A number of studies have examined the structural properties of late folding intermediates of (β/α)8-barrel proteins involved in tryptophan biosynthesis, whereas there is little information available about the early folding events of these proteins. To identify the contiguous polypeptide segments important to the folding of the (β/α)8-barrel protein Escherichia coli N-(5′-phosphoribosyl)anthranilate isomerase, we structurally characterized fragments and circularly permuted forms of the protein. We also simulated thermal unfolding of the protein using molecular dynamics. Our fragmentation experiments demonstrate that the isolated (β/α)1-4β5 fragment is almost as stable as the full-length protein. The far and near-UV CD spectra of this fragment are indicative of native-like secondary and tertiary structures. Structural analysis of the circularly permutated proteins shows that if the protein is cleaved within the two N-terminal βα modules, the amount of secondary structure is unaffected, whereas, when cleaved within the central (β/α)3-4β5 segment, the protein simply cannot fold. An ensemble of the denatured structures produced by thermal unfolding simulations contains a persistent local structure comprised of β3, β4 and β5. The presence of this three-stranded β-barrel suggests that it may be an important early-stage folding intermediate. Interactions found in (β/α) 3-4β5 may be essential for the early events of ePRAI folding if they provide a nucleation site that directs folding.
AB - A number of studies have examined the structural properties of late folding intermediates of (β/α)8-barrel proteins involved in tryptophan biosynthesis, whereas there is little information available about the early folding events of these proteins. To identify the contiguous polypeptide segments important to the folding of the (β/α)8-barrel protein Escherichia coli N-(5′-phosphoribosyl)anthranilate isomerase, we structurally characterized fragments and circularly permuted forms of the protein. We also simulated thermal unfolding of the protein using molecular dynamics. Our fragmentation experiments demonstrate that the isolated (β/α)1-4β5 fragment is almost as stable as the full-length protein. The far and near-UV CD spectra of this fragment are indicative of native-like secondary and tertiary structures. Structural analysis of the circularly permutated proteins shows that if the protein is cleaved within the two N-terminal βα modules, the amount of secondary structure is unaffected, whereas, when cleaved within the central (β/α)3-4β5 segment, the protein simply cannot fold. An ensemble of the denatured structures produced by thermal unfolding simulations contains a persistent local structure comprised of β3, β4 and β5. The presence of this three-stranded β-barrel suggests that it may be an important early-stage folding intermediate. Interactions found in (β/α) 3-4β5 may be essential for the early events of ePRAI folding if they provide a nucleation site that directs folding.
KW - (β/α)-barrel protein
KW - Circular permutation
KW - Fragmentation
KW - Molecular dynamics simulation
KW - Protein folding
UR - http://www.scopus.com/inward/record.url?scp=27144517647&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=27144517647&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2005.08.070
DO - 10.1016/j.jmb.2005.08.070
M3 - Article
C2 - 16216267
AN - SCOPUS:27144517647
VL - 353
SP - 1161
EP - 1170
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 5
ER -