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Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein

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Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein. / Löw, Christian; Weininger, Ulrich; Neumann, Piotr; Klepsch, Mirjam; Lilie, Hauke; Stubbs, Milton T; Balbach, Jochen.

in: P NATL ACAD SCI USA, Jahrgang 105, Nr. 10, 11.03.2008, S. 3779-84.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Löw, C, Weininger, U, Neumann, P, Klepsch, M, Lilie, H, Stubbs, MT & Balbach, J 2008, 'Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein', P NATL ACAD SCI USA, Jg. 105, Nr. 10, S. 3779-84. https://doi.org/10.1073/pnas.0710657105

APA

Löw, C., Weininger, U., Neumann, P., Klepsch, M., Lilie, H., Stubbs, M. T., & Balbach, J. (2008). Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein. P NATL ACAD SCI USA, 105(10), 3779-84. https://doi.org/10.1073/pnas.0710657105

Vancouver

Löw C, Weininger U, Neumann P, Klepsch M, Lilie H, Stubbs MT et al. Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein. P NATL ACAD SCI USA. 2008 Mär 11;105(10):3779-84. https://doi.org/10.1073/pnas.0710657105

Bibtex

@article{6952365f72504c3d90558772009c4c3a,
title = "Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein",
abstract = "Repeat proteins are widespread in nature, with many of them functioning as binding molecules in protein-protein recognition. Their simple structural architecture is used in biotechnology for generating proteins with high affinities to target proteins. Recent folding studies of ankyrin repeat (AR) proteins revealed a new mechanism of protein folding. The formation of an intermediate state is rate limiting in the folding reaction, suggesting a scaffold function of this transient state for intrinsically less stable ARs. To investigate a possible common mechanism of AR folding, we studied the structure and folding of a new thermophilic AR protein (tANK) identified in the archaeon Thermoplasma volcanium. The x-ray structure of the evolutionary much older tANK revealed high homology to the human CDK inhibitor p19(INK4d), whose sequence was used for homology search. As for p19(INK4d), equilibrium and kinetic folding analyses classify tANK to the family of sequential three-state folding proteins, with an unusual fast equilibrium between native and intermediate state. Under equilibrium conditions, the intermediate can be populated to >90%, allowing characterization on a residue-by-residue level using NMR spectroscopy. These data clearly show that the three C-terminal ARs are natively folded in the intermediate state, whereas native cross-peaks for the rest of the molecule are missing. Therefore, the formation of a stable folding unit consisting of three ARs is the necessary rate-limiting step before AR 1 and 2 can assemble to form the native state.",
keywords = "Ankyrin Repeat, Archaeal Proteins, Circular Dichroism, Crystallography, X-Ray, Guanidine, Humans, Kinetics, Models, Molecular, Nitrogen Isotopes, Nuclear Magnetic Resonance, Biomolecular, Protein Denaturation, Protein Folding, Spectrometry, Fluorescence, Thermoplasma, Journal Article, Research Support, Non-U.S. Gov't",
author = "Christian L{\"o}w and Ulrich Weininger and Piotr Neumann and Mirjam Klepsch and Hauke Lilie and Stubbs, {Milton T} and Jochen Balbach",
year = "2008",
month = mar,
day = "11",
doi = "10.1073/pnas.0710657105",
language = "English",
volume = "105",
pages = "3779--84",
journal = "P NATL ACAD SCI USA",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "10",

}

RIS

TY - JOUR

T1 - Structural insights into an equilibrium folding intermediate of an archaeal ankyrin repeat protein

AU - Löw, Christian

AU - Weininger, Ulrich

AU - Neumann, Piotr

AU - Klepsch, Mirjam

AU - Lilie, Hauke

AU - Stubbs, Milton T

AU - Balbach, Jochen

PY - 2008/3/11

Y1 - 2008/3/11

N2 - Repeat proteins are widespread in nature, with many of them functioning as binding molecules in protein-protein recognition. Their simple structural architecture is used in biotechnology for generating proteins with high affinities to target proteins. Recent folding studies of ankyrin repeat (AR) proteins revealed a new mechanism of protein folding. The formation of an intermediate state is rate limiting in the folding reaction, suggesting a scaffold function of this transient state for intrinsically less stable ARs. To investigate a possible common mechanism of AR folding, we studied the structure and folding of a new thermophilic AR protein (tANK) identified in the archaeon Thermoplasma volcanium. The x-ray structure of the evolutionary much older tANK revealed high homology to the human CDK inhibitor p19(INK4d), whose sequence was used for homology search. As for p19(INK4d), equilibrium and kinetic folding analyses classify tANK to the family of sequential three-state folding proteins, with an unusual fast equilibrium between native and intermediate state. Under equilibrium conditions, the intermediate can be populated to >90%, allowing characterization on a residue-by-residue level using NMR spectroscopy. These data clearly show that the three C-terminal ARs are natively folded in the intermediate state, whereas native cross-peaks for the rest of the molecule are missing. Therefore, the formation of a stable folding unit consisting of three ARs is the necessary rate-limiting step before AR 1 and 2 can assemble to form the native state.

AB - Repeat proteins are widespread in nature, with many of them functioning as binding molecules in protein-protein recognition. Their simple structural architecture is used in biotechnology for generating proteins with high affinities to target proteins. Recent folding studies of ankyrin repeat (AR) proteins revealed a new mechanism of protein folding. The formation of an intermediate state is rate limiting in the folding reaction, suggesting a scaffold function of this transient state for intrinsically less stable ARs. To investigate a possible common mechanism of AR folding, we studied the structure and folding of a new thermophilic AR protein (tANK) identified in the archaeon Thermoplasma volcanium. The x-ray structure of the evolutionary much older tANK revealed high homology to the human CDK inhibitor p19(INK4d), whose sequence was used for homology search. As for p19(INK4d), equilibrium and kinetic folding analyses classify tANK to the family of sequential three-state folding proteins, with an unusual fast equilibrium between native and intermediate state. Under equilibrium conditions, the intermediate can be populated to >90%, allowing characterization on a residue-by-residue level using NMR spectroscopy. These data clearly show that the three C-terminal ARs are natively folded in the intermediate state, whereas native cross-peaks for the rest of the molecule are missing. Therefore, the formation of a stable folding unit consisting of three ARs is the necessary rate-limiting step before AR 1 and 2 can assemble to form the native state.

KW - Ankyrin Repeat

KW - Archaeal Proteins

KW - Circular Dichroism

KW - Crystallography, X-Ray

KW - Guanidine

KW - Humans

KW - Kinetics

KW - Models, Molecular

KW - Nitrogen Isotopes

KW - Nuclear Magnetic Resonance, Biomolecular

KW - Protein Denaturation

KW - Protein Folding

KW - Spectrometry, Fluorescence

KW - Thermoplasma

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1073/pnas.0710657105

DO - 10.1073/pnas.0710657105

M3 - SCORING: Journal article

C2 - 18305166

VL - 105

SP - 3779

EP - 3784

JO - P NATL ACAD SCI USA

JF - P NATL ACAD SCI USA

SN - 0027-8424

IS - 10

ER -