Research ExplorerPublicationsTryptophan Scanning Mutagenesis of EF-Hand Motifs

Tryptophan Scanning Mutagenesis of EF-Hand Motifs

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Tryptophan Scanning Mutagenesis of EF-Hand Motifs. / Kiran, Uday; Kreutz, Michael R; Sharma, Yogendra; Chakraborty, Asima.

In: Methods Mol Biol, Vol. 1929, 02.02.2019, p. 567-581.

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

Harvard

Kiran, U, Kreutz, MR, Sharma, Y & Chakraborty, A 2019, 'Tryptophan Scanning Mutagenesis of EF-Hand Motifs', Methods Mol Biol, vol. 1929, pp. 567-581. https://doi.org/10.1007/978-1-4939-9030-6_35

APA

Kiran, U., Kreutz, M. R., Sharma, Y., & Chakraborty, A. (2019). Tryptophan Scanning Mutagenesis of EF-Hand Motifs. Methods Mol Biol, 1929, 567-581. https://doi.org/10.1007/978-1-4939-9030-6_35

Vancouver

Kiran U, Kreutz MR, Sharma Y, Chakraborty A. Tryptophan Scanning Mutagenesis of EF-Hand Motifs. Methods Mol Biol. 2019 Feb 2;1929:567-581. https://doi.org/10.1007/978-1-4939-9030-6_35

Bibtex

@article{fccc2cd5861744aa9d99913d36c1108d,
title = "Tryptophan Scanning Mutagenesis of EF-Hand Motifs",
abstract = "Ca2+ regulation in living systems occurs via specific structural alterations, subtle or drastic, in the Ca2+-binding domains of sensor proteins. Sensor proteins perform designated nonredundant roles within the dense network of Ca2+-binding proteins. A detailed understanding of the structural changes in calcium sensor proteins due to Ca2+ spikes that vary spatially, temporally, and in magnitude would provide better insights into the mechanism of Ca2+ sensing. This chapter describes a method to study various stages during apo to the holo transition of Ca2+-binding proteins by Trp-mediated scanning of individual EF-hand motifs. We describe the applicability of this procedure to caldendrin, which is a neuronal Ca2+-binding protein and to integrin-binding protein. Tryptophan mutants of full-length caldendrin were designed to reveal local structural changes in each EF-hand of the protein. This method, referred to as {"}EF-hand scanning tryptophan mutagenesis,{"} not only allows the identification of canonical and noncanonical EF-hands using very low concentrations of protein but also enables visualization of the hierarchical filling of Ca2+ into the canonical EF-hands.",
keywords = "Journal Article, Research Support, Non-U.S. Gov't",
author = "Uday Kiran and Kreutz, {Michael R} and Yogendra Sharma and Asima Chakraborty",
year = "2019",
month = feb,
day = "2",
doi = "10.1007/978-1-4939-9030-6_35",
language = "English",
volume = "1929",
pages = "567--581",
journal = "Methods Mol Biol",
issn = "1064-3745",
publisher = "Humana Press",

}

RIS

TY - JOUR

T1 - Tryptophan Scanning Mutagenesis of EF-Hand Motifs

AU - Kiran, Uday

AU - Kreutz, Michael R

AU - Sharma, Yogendra

AU - Chakraborty, Asima

PY - 2019/2/2

Y1 - 2019/2/2

N2 - Ca2+ regulation in living systems occurs via specific structural alterations, subtle or drastic, in the Ca2+-binding domains of sensor proteins. Sensor proteins perform designated nonredundant roles within the dense network of Ca2+-binding proteins. A detailed understanding of the structural changes in calcium sensor proteins due to Ca2+ spikes that vary spatially, temporally, and in magnitude would provide better insights into the mechanism of Ca2+ sensing. This chapter describes a method to study various stages during apo to the holo transition of Ca2+-binding proteins by Trp-mediated scanning of individual EF-hand motifs. We describe the applicability of this procedure to caldendrin, which is a neuronal Ca2+-binding protein and to integrin-binding protein. Tryptophan mutants of full-length caldendrin were designed to reveal local structural changes in each EF-hand of the protein. This method, referred to as "EF-hand scanning tryptophan mutagenesis," not only allows the identification of canonical and noncanonical EF-hands using very low concentrations of protein but also enables visualization of the hierarchical filling of Ca2+ into the canonical EF-hands.

AB - Ca2+ regulation in living systems occurs via specific structural alterations, subtle or drastic, in the Ca2+-binding domains of sensor proteins. Sensor proteins perform designated nonredundant roles within the dense network of Ca2+-binding proteins. A detailed understanding of the structural changes in calcium sensor proteins due to Ca2+ spikes that vary spatially, temporally, and in magnitude would provide better insights into the mechanism of Ca2+ sensing. This chapter describes a method to study various stages during apo to the holo transition of Ca2+-binding proteins by Trp-mediated scanning of individual EF-hand motifs. We describe the applicability of this procedure to caldendrin, which is a neuronal Ca2+-binding protein and to integrin-binding protein. Tryptophan mutants of full-length caldendrin were designed to reveal local structural changes in each EF-hand of the protein. This method, referred to as "EF-hand scanning tryptophan mutagenesis," not only allows the identification of canonical and noncanonical EF-hands using very low concentrations of protein but also enables visualization of the hierarchical filling of Ca2+ into the canonical EF-hands.

KW - Journal Article

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

U2 - 10.1007/978-1-4939-9030-6_35

DO - 10.1007/978-1-4939-9030-6_35

M3 - SCORING: Journal article

C2 - 30710297

VL - 1929

SP - 567

EP - 581

JO - Methods Mol Biol

JF - Methods Mol Biol

SN - 1064-3745

ER -