Azidoethoxyphenylalanine as a Vibrational Reporter and Click Chemistry Partner in Proteins

Tookmanian, Elise M.; Phillips-Piro, Christine M.; Fenlon, Edward E.; Brewer, Scott H.

doi:10.1002/chem.201503908

Title: Azidoethoxyphenylalanine as a Vibrational Reporter and Click Chemistry Partner in Proteins

Journal Article · Thu Nov 26 00:00:00 EST 2015 · Chemistry - A European Journal

DOI:https://doi.org/10.1002/chem.201503908· OSTI ID:1237757

Tookmanian, Elise M. ^[1]; Phillips-Piro, Christine M. ^[1]; Fenlon, Edward E. ^[1]; Brewer, Scott H. ^[1]

Franklin & Marshall College, Lancaster, PA (United States)

Abstract An unnatural amino acid, 4‐(2‐azidoethoxy)‐ L ‐phenylalanine (AePhe, 1 ), was designed and synthesized in three steps from known compounds in 54 % overall yield. The sensitivity of the IR absorption of the azide of AePhe was established by comparison of the frequency of the azide asymmetric stretch vibration in water and dimethyl sulfoxide. AePhe was successfully incorporated into superfolder green fluorescent protein (sfGFP) at the 133 and 149 sites by using the amber codon suppression method. The IR spectra of these sfGFP constructs indicated that the azide group at the 149 site was not fully solvated despite the location in sfGFP and the three‐atom linker between the azido group and the aromatic ring of AePhe. An X‐ray crystal structure of sfGFP‐149‐AePhe was solved at 1.45 Å resolution and provides an explanation for the IR data as the flexible linker adopts a conformation which partially buries the azide on the protein surface. Both sfGFP‐AePhe constructs efficiently undergo a bioorthogonal strain‐promoted click cycloaddition with a dibenzocyclooctyne derivative.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC); National Science Foundation (NSF); NIH NIGMS

Grant/Contract Number:: AC02‐06CH11357; CHE‐1053946; P41 GM103403; AC02-06CH11357

OSTI ID:: 1237757

Alternate ID(s):: OSTI ID: 1786149

Journal Information:: Chemistry - A European Journal, Vol. 21, Issue 52; ISSN 0947-6539

Publisher:: ChemPubSoc EuropeCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 10 works

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