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Daniel Otzen

Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway

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Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway. / De Prat Gay, Gonzalo; Ruiz-Sanz, Javier; Neira, José L.; Corrales, Fernando J.; Otzen, Daniel E.; Ladurner, Andreas G.; Fersht, Alan R.

In: Journal of Molecular Biology, Vol. 254, No. 5, 15.12.1995, p. 968-979.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

De Prat Gay, G, Ruiz-Sanz, J, Neira, JL, Corrales, FJ, Otzen, DE, Ladurner, AG & Fersht, AR 1995, 'Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway', Journal of Molecular Biology, vol. 254, no. 5, pp. 968-979. https://doi.org/10.1006/jmbi.1995.0669

APA

De Prat Gay, G., Ruiz-Sanz, J., Neira, J. L., Corrales, F. J., Otzen, D. E., Ladurner, A. G., & Fersht, A. R. (1995). Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway. Journal of Molecular Biology, 254(5), 968-979. https://doi.org/10.1006/jmbi.1995.0669

CBE

MLA

Vancouver

Author

De Prat Gay, Gonzalo ; Ruiz-Sanz, Javier ; Neira, José L. ; Corrales, Fernando J. ; Otzen, Daniel E. ; Ladurner, Andreas G. ; Fersht, Alan R. / Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway. In: Journal of Molecular Biology. 1995 ; Vol. 254, No. 5. pp. 968-979.

Bibtex

@article{38932d0c4db74452ad28360a202402fc,
title = "Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway",
abstract = "We have obtained a series of fragments growing from the N terminus of the protein chymotrypsin inhibitor-2 (CI2) in order to study the development of structure on elongation of the polypeptide in solution. We present an extensive biophysical characterization of ten fragments using different conformational probes. Small fragments up to residue 40 of the 64-residue protein are disordered. Fragment (1-40) has non-native local hydrophobic clusters, but nevertheless does not bind 8-anilinonaphthalene-1-sulphonate (ANS). Hydrophobic regions in longer fragments become gradually more capable of binding ANS as the chain grows to completion, with a tendency to form native structures. Major changes in secondary structure and accessibility to hydrophobic sites occur in parallel, between (1-40) and (1-53), together with changes in hydrodynamic volume and flexibility NMR studies of (1-53), the first fragment displaying tertiary interactions, show that a subcore is fully formed and the α-helix (residues 12 to 24) is of fluctuating structure. Fragments (1-53) and (1-60) share many properties with molten globule-like structures, with varying degrees or order. Fluorescence properties of the native fold are gradually recovered from fragments (1-60) to full-length CI2, together with a decrease in hydrophobic exposure. A small degree of co-operativity of formation of structure appears when residue 60 is added, gradually increasing as residue 62 is added, but a full two-state co-operative transition appears only on addition of Arg62 and Val63. We believe this is the result of correct side-chain packing of the hydrophobic core, capping the major elements of secondary structure in CI2 at this late stage, which is probed by the complete recovery of the fluorescence of the unique Trp5. The structures that develop as the polypeptide chain increases in length parallel the structural features present in the nucleus for the folding of intact protein, which develops in the transition state. The folding nucleus consists of much of the helix and the interactions made by Ala16 in the helix with residues in the core, especially with Leu49 and Ile57, with the rest of the structure being formed only very weakly in the transition state.",
keywords = "CI2, Cooperativity, Nascent, Polypeptide, Protein folding",
author = "{De Prat Gay}, Gonzalo and Javier Ruiz-Sanz and Neira, {Jos{\'e} L.} and Corrales, {Fernando J.} and Otzen, {Daniel E.} and Ladurner, {Andreas G.} and Fersht, {Alan R.}",
year = "1995",
month = dec,
day = "15",
doi = "10.1006/jmbi.1995.0669",
language = "English",
volume = "254",
pages = "968--979",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press",
number = "5",

}

RIS

TY - JOUR

T1 - Conformational pathway of the polypeptide chain of chymotrypsin inhibitor-2 growing from its N terminus in vitro. Parallels with the protein folding pathway

AU - De Prat Gay, Gonzalo

AU - Ruiz-Sanz, Javier

AU - Neira, José L.

AU - Corrales, Fernando J.

AU - Otzen, Daniel E.

AU - Ladurner, Andreas G.

AU - Fersht, Alan R.

PY - 1995/12/15

Y1 - 1995/12/15

N2 - We have obtained a series of fragments growing from the N terminus of the protein chymotrypsin inhibitor-2 (CI2) in order to study the development of structure on elongation of the polypeptide in solution. We present an extensive biophysical characterization of ten fragments using different conformational probes. Small fragments up to residue 40 of the 64-residue protein are disordered. Fragment (1-40) has non-native local hydrophobic clusters, but nevertheless does not bind 8-anilinonaphthalene-1-sulphonate (ANS). Hydrophobic regions in longer fragments become gradually more capable of binding ANS as the chain grows to completion, with a tendency to form native structures. Major changes in secondary structure and accessibility to hydrophobic sites occur in parallel, between (1-40) and (1-53), together with changes in hydrodynamic volume and flexibility NMR studies of (1-53), the first fragment displaying tertiary interactions, show that a subcore is fully formed and the α-helix (residues 12 to 24) is of fluctuating structure. Fragments (1-53) and (1-60) share many properties with molten globule-like structures, with varying degrees or order. Fluorescence properties of the native fold are gradually recovered from fragments (1-60) to full-length CI2, together with a decrease in hydrophobic exposure. A small degree of co-operativity of formation of structure appears when residue 60 is added, gradually increasing as residue 62 is added, but a full two-state co-operative transition appears only on addition of Arg62 and Val63. We believe this is the result of correct side-chain packing of the hydrophobic core, capping the major elements of secondary structure in CI2 at this late stage, which is probed by the complete recovery of the fluorescence of the unique Trp5. The structures that develop as the polypeptide chain increases in length parallel the structural features present in the nucleus for the folding of intact protein, which develops in the transition state. The folding nucleus consists of much of the helix and the interactions made by Ala16 in the helix with residues in the core, especially with Leu49 and Ile57, with the rest of the structure being formed only very weakly in the transition state.

AB - We have obtained a series of fragments growing from the N terminus of the protein chymotrypsin inhibitor-2 (CI2) in order to study the development of structure on elongation of the polypeptide in solution. We present an extensive biophysical characterization of ten fragments using different conformational probes. Small fragments up to residue 40 of the 64-residue protein are disordered. Fragment (1-40) has non-native local hydrophobic clusters, but nevertheless does not bind 8-anilinonaphthalene-1-sulphonate (ANS). Hydrophobic regions in longer fragments become gradually more capable of binding ANS as the chain grows to completion, with a tendency to form native structures. Major changes in secondary structure and accessibility to hydrophobic sites occur in parallel, between (1-40) and (1-53), together with changes in hydrodynamic volume and flexibility NMR studies of (1-53), the first fragment displaying tertiary interactions, show that a subcore is fully formed and the α-helix (residues 12 to 24) is of fluctuating structure. Fragments (1-53) and (1-60) share many properties with molten globule-like structures, with varying degrees or order. Fluorescence properties of the native fold are gradually recovered from fragments (1-60) to full-length CI2, together with a decrease in hydrophobic exposure. A small degree of co-operativity of formation of structure appears when residue 60 is added, gradually increasing as residue 62 is added, but a full two-state co-operative transition appears only on addition of Arg62 and Val63. We believe this is the result of correct side-chain packing of the hydrophobic core, capping the major elements of secondary structure in CI2 at this late stage, which is probed by the complete recovery of the fluorescence of the unique Trp5. The structures that develop as the polypeptide chain increases in length parallel the structural features present in the nucleus for the folding of intact protein, which develops in the transition state. The folding nucleus consists of much of the helix and the interactions made by Ala16 in the helix with residues in the core, especially with Leu49 and Ile57, with the rest of the structure being formed only very weakly in the transition state.

KW - CI2

KW - Cooperativity

KW - Nascent

KW - Polypeptide

KW - Protein folding

UR - http://www.scopus.com/inward/record.url?scp=0029558960&partnerID=8YFLogxK

U2 - 10.1006/jmbi.1995.0669

DO - 10.1006/jmbi.1995.0669

M3 - Journal article

C2 - 7500364

AN - SCOPUS:0029558960

VL - 254

SP - 968

EP - 979

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 5

ER -