A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede

Kwan Ho Jung, Matthew Fares, Leeann S. Grainger, Charles H. Wolstenholme, Anna Hou, Yu Liu, Xin Zhang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Self-labelling protein tags with fluorogenic probes serve as great fluorescence imaging tools to understand key questions of protein dynamics and functions in living cells. In the present study, we report a SNAP-tag fluorogenic probe 4c mimicking the chromophore of the red fluorescent protein Kaede. The molecular rotor properties of 4c were utilized as a fluorogenic probe for SNAP-tag, such that conjugation with SNAPf protein leads to inhibition of twisted intramolecular charge transfer, triggering fluorogenecity. Upon conjugation with SNAPf, 4c exhibited approximately a 90-fold enhancement in fluorescence intensity with fast labelling kinetics (k 2 = 15000 M -1 s -1 ). Biochemical and spectroscopic studies confirmed that fluorescence requires formation of folded SNAPf·4c covalent conjugate between Cys 145 and 4c. Confocal microscopy and flow cytometry showed that 4c is capable of detecting SNAPf proteins or SNAPf fused with a protein of interest in living cells. This work provides a framework to develop the large family of FP chromophores into fluorogenic probes for self-labelling protein tags.

Original languageEnglish (US)
Pages (from-to)1906-1915
Number of pages10
JournalOrganic and Biomolecular Chemistry
Volume17
Issue number7
DOIs
StatePublished - Jan 1 2019

Fingerprint

SNAP
Chromophores
chromophores
proteins
probes
Labeling
Proteins
marking
Fluorescence
conjugation
fluorescence
Cells
Flow cytometry
Confocal microscopy
Optical Imaging
cytometry
Confocal Microscopy
red fluorescent protein
Charge transfer
Flow Cytometry

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physical and Theoretical Chemistry
  • Organic Chemistry

Cite this

Jung, K. H., Fares, M., Grainger, L. S., Wolstenholme, C. H., Hou, A., Liu, Y., & Zhang, X. (2019). A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede. Organic and Biomolecular Chemistry, 17(7), 1906-1915. https://doi.org/10.1039/c8ob01483c
Jung, Kwan Ho ; Fares, Matthew ; Grainger, Leeann S. ; Wolstenholme, Charles H. ; Hou, Anna ; Liu, Yu ; Zhang, Xin. / A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede. In: Organic and Biomolecular Chemistry. 2019 ; Vol. 17, No. 7. pp. 1906-1915.
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abstract = "Self-labelling protein tags with fluorogenic probes serve as great fluorescence imaging tools to understand key questions of protein dynamics and functions in living cells. In the present study, we report a SNAP-tag fluorogenic probe 4c mimicking the chromophore of the red fluorescent protein Kaede. The molecular rotor properties of 4c were utilized as a fluorogenic probe for SNAP-tag, such that conjugation with SNAPf protein leads to inhibition of twisted intramolecular charge transfer, triggering fluorogenecity. Upon conjugation with SNAPf, 4c exhibited approximately a 90-fold enhancement in fluorescence intensity with fast labelling kinetics (k 2 = 15000 M -1 s -1 ). Biochemical and spectroscopic studies confirmed that fluorescence requires formation of folded SNAPf·4c covalent conjugate between Cys 145 and 4c. Confocal microscopy and flow cytometry showed that 4c is capable of detecting SNAPf proteins or SNAPf fused with a protein of interest in living cells. This work provides a framework to develop the large family of FP chromophores into fluorogenic probes for self-labelling protein tags.",
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Jung, KH, Fares, M, Grainger, LS, Wolstenholme, CH, Hou, A, Liu, Y & Zhang, X 2019, 'A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede', Organic and Biomolecular Chemistry, vol. 17, no. 7, pp. 1906-1915. https://doi.org/10.1039/c8ob01483c

A SNAP-tag fluorogenic probe mimicking the chromophore of the red fluorescent protein Kaede. / Jung, Kwan Ho; Fares, Matthew; Grainger, Leeann S.; Wolstenholme, Charles H.; Hou, Anna; Liu, Yu; Zhang, Xin.

In: Organic and Biomolecular Chemistry, Vol. 17, No. 7, 01.01.2019, p. 1906-1915.

Research output: Contribution to journalArticle

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AU - Fares, Matthew

AU - Grainger, Leeann S.

AU - Wolstenholme, Charles H.

AU - Hou, Anna

AU - Liu, Yu

AU - Zhang, Xin

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