Fluorescent proteins: Crystallization, structural determination, and nonnatural amino acid incorporation

Ahmed, Rochelle D., Auhim, Husam Sabah, Worthy, Harley L. and Jones, D. Dafydd ORCID: https://orcid.org/0000-0001-7709-3995 2023. Fluorescent proteins: Crystallization, structural determination, and nonnatural amino acid incorporation. Sharma, Mayank, ed. Fluorescent Proteins: Methods and Protocols, Vol. 2564. Methods in Molecular Biology, Springer, pp. 99-119. (10.1007/978-1-0716-2667-2_5)

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Abstract

Fluorescent proteins have revolutionized cell biology and cell imaging through their use as genetically encoded tags. Structural biology has been pivotal in understanding how their unique fluorescent properties manifest through the formation of the chromophore and how the spectral properties are tuned through interaction networks. This knowledge has in turn led to the construction of novel variants with new and improved properties. Here we describe the process by which fluorescent protein structures are determined, starting from recombinant protein production to structure determination by molecular replacement. We also describe how to incorporate and determine the structures of proteins containing non-natural amino acids. Recent advances in protein engineering have led to reprogramming of the genetic code to allow incorporation of new chemistry at designed residue positions, with fluorescent proteins being at the forefront of structural studies in this area. The impact of such new chemistry on protein structure is still limited; the accumulation of more protein structures will undoubtedly improve our understanding and ability to engineer proteins with new chemical functionality.

Item Type:	Book Section
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Publisher:	Springer
ISBN:	9781071626672
ISSN:	1064-3745
Date of First Compliant Deposit:	19 October 2022
Date of Acceptance:	1 September 2022
Last Modified:	06 Jan 2024 04:20
URI:	https://orca.cardiff.ac.uk/id/eprint/153512

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