Slow, reversible, coupled folding and binding of the spectrin tetramerization domain

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Slow, reversible, coupled folding and binding of the spectrin tetramerization domain. / Shammas, S. L.; Rogers, J. M.; Hill, S. A.; Clarke, J.

In: Biophysical Journal, Vol. 103, No. 10, 21.11.2012, p. 2203-2214.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Shammas, SL, Rogers, JM, Hill, SA & Clarke, J 2012, 'Slow, reversible, coupled folding and binding of the spectrin tetramerization domain', Biophysical Journal, vol. 103, no. 10, pp. 2203-2214. https://doi.org/10.1016/j.bpj.2012.10.012

APA

Shammas, S. L., Rogers, J. M., Hill, S. A., & Clarke, J. (2012). Slow, reversible, coupled folding and binding of the spectrin tetramerization domain. Biophysical Journal, 103(10), 2203-2214. https://doi.org/10.1016/j.bpj.2012.10.012

Vancouver

Shammas SL, Rogers JM, Hill SA, Clarke J. Slow, reversible, coupled folding and binding of the spectrin tetramerization domain. Biophysical Journal. 2012 Nov 21;103(10):2203-2214. https://doi.org/10.1016/j.bpj.2012.10.012

Author

Shammas, S. L. ; Rogers, J. M. ; Hill, S. A. ; Clarke, J. / Slow, reversible, coupled folding and binding of the spectrin tetramerization domain. In: Biophysical Journal. 2012 ; Vol. 103, No. 10. pp. 2203-2214.

Bibtex

@article{5d0d3c13272345cbb71789748af8210a,
title = "Slow, reversible, coupled folding and binding of the spectrin tetramerization domain",
abstract = "Many intrinsically disordered proteins (IDPs) are significantly unstructured under physiological conditions. A number of these IDPs have been shown to undergo coupled folding and binding reactions whereby they can gain structure upon association with an appropriate partner protein. In general, these systems display weaker binding affinities than do systems with association between completely structured domains, with micromolar Kd values appearing typical. One such system is the association between α- and β-spectrin, where two partially structured, incomplete domains associate to form a fully structured, three-helix bundle, the spectrin tetramerization domain. Here, we use this model system to demonstrate a method for fitting association and dissociation kinetic traces where, using typical biophysical concentrations, the association reactions are expected to be highly reversible. We elucidate the unusually slow, two-state kinetics of spectrin assembly in solution. The advantages of studying kinetics in this regime include the potential for gaining equilibrium constants as well as rate constants, and for performing experiments with low protein concentrations. We suggest that this approach would be particularly appropriate for high-throughput mutational analysis of two-state reversible binding processes.",
author = "Shammas, {S. L.} and Rogers, {J. M.} and Hill, {S. A.} and J. Clarke",
year = "2012",
month = nov,
day = "21",
doi = "10.1016/j.bpj.2012.10.012",
language = "English",
volume = "103",
pages = "2203--2214",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Cell Press",
number = "10",

}

RIS

TY - JOUR

T1 - Slow, reversible, coupled folding and binding of the spectrin tetramerization domain

AU - Shammas, S. L.

AU - Rogers, J. M.

AU - Hill, S. A.

AU - Clarke, J.

PY - 2012/11/21

Y1 - 2012/11/21

N2 - Many intrinsically disordered proteins (IDPs) are significantly unstructured under physiological conditions. A number of these IDPs have been shown to undergo coupled folding and binding reactions whereby they can gain structure upon association with an appropriate partner protein. In general, these systems display weaker binding affinities than do systems with association between completely structured domains, with micromolar Kd values appearing typical. One such system is the association between α- and β-spectrin, where two partially structured, incomplete domains associate to form a fully structured, three-helix bundle, the spectrin tetramerization domain. Here, we use this model system to demonstrate a method for fitting association and dissociation kinetic traces where, using typical biophysical concentrations, the association reactions are expected to be highly reversible. We elucidate the unusually slow, two-state kinetics of spectrin assembly in solution. The advantages of studying kinetics in this regime include the potential for gaining equilibrium constants as well as rate constants, and for performing experiments with low protein concentrations. We suggest that this approach would be particularly appropriate for high-throughput mutational analysis of two-state reversible binding processes.

AB - Many intrinsically disordered proteins (IDPs) are significantly unstructured under physiological conditions. A number of these IDPs have been shown to undergo coupled folding and binding reactions whereby they can gain structure upon association with an appropriate partner protein. In general, these systems display weaker binding affinities than do systems with association between completely structured domains, with micromolar Kd values appearing typical. One such system is the association between α- and β-spectrin, where two partially structured, incomplete domains associate to form a fully structured, three-helix bundle, the spectrin tetramerization domain. Here, we use this model system to demonstrate a method for fitting association and dissociation kinetic traces where, using typical biophysical concentrations, the association reactions are expected to be highly reversible. We elucidate the unusually slow, two-state kinetics of spectrin assembly in solution. The advantages of studying kinetics in this regime include the potential for gaining equilibrium constants as well as rate constants, and for performing experiments with low protein concentrations. We suggest that this approach would be particularly appropriate for high-throughput mutational analysis of two-state reversible binding processes.

UR - http://www.scopus.com/inward/record.url?scp=84869387199&partnerID=8YFLogxK

U2 - 10.1016/j.bpj.2012.10.012

DO - 10.1016/j.bpj.2012.10.012

M3 - Journal article

C2 - 23200054

AN - SCOPUS:84869387199

VL - 103

SP - 2203

EP - 2214

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 10

ER -

ID: 244651260