Sensitive, site-specific, and stable vibrational probe of local protein environments: 4-azidomethyl-L-phenylalanine.
Publication/Presentation Date
8-1-2013
Abstract
We have synthesized the unnatural amino acid (UAA), 4-azidomethyl-L-phenylalanine (pN₃CH₂Phe), to serve as an effective vibrational reporter of local protein environments. The position, extinction coefficient, and sensitivity to local environment of the azide asymmetric stretch vibration of pN₃CH₂Phe are compared to the vibrational reporters: 4-cyano-L-phenylalanine (pCNPhe) and 4-azido-L-phenylalanine (pN₃Phe). This UAA was genetically incorporated in a site-specific manner utilizing an engineered, orthogonal aminoacyl-tRNA synthetase in response to an amber codon with high efficiency and fidelity into two distinct sites in superfolder green fluorescent protein (sfGFP). This allowed for the dependence of the azide asymmetric stretch vibration of pN₃CH₂Phe to different protein environments to be measured. The photostability of pN₃CH₂Phe was also measured relative to the photoreactive UAA, pN₃Phe.
Volume
117
Issue
30
First Page
8987
Last Page
8993
ISSN
1520-5207
Published In/Presented At
Bazewicz, C. G., Liskov, M. T., Hines, K. J., & Brewer, S. H. (2013). Sensitive, site-specific, and stable vibrational probe of local protein environments: 4-azidomethyl-L-phenylalanine. The journal of physical chemistry. B, 117(30), 8987–8993. https://doi.org/10.1021/jp4052598
Disciplines
Medicine and Health Sciences
PubMedID
23865850
Department(s)
Department of Surgery
Document Type
Article
