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Angew Chem Int Ed Engl. 2015 May 4;54(19):5784-8. doi: 10.1002/anie.201411692. Epub 2015 Mar 17.

Secondary-ion mass spectrometry of genetically encoded targets.

Author information

1
Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, Humboldtallee 23, 37073 Göttingen (Germany); Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen (Germany); International Max Planck Research School Molecular Biology, Göttingen (Germany).

Abstract

Secondary ion mass spectrometry (SIMS) is generally used in imaging the isotopic composition of various materials. It is becoming increasingly popular in biology, especially for investigations of cellular metabolism. However, individual proteins are difficult to identify in SIMS, which limits the ability of this technology to study individual compartments or protein complexes. We present a method for specific protein isotopic and fluorescence labeling (SPILL), based on a novel click reaction with isotopic probes. Using this method, we added (19) F-enriched labels to different proteins, and visualized them by NanoSIMS and fluorescence microscopy. The (19) F signal allowed the precise visualization of the protein of interest, with minimal background, and enabled correlative studies of protein distribution and cellular metabolism or composition. SPILL can be applied to biological systems suitable for click chemistry, which include most cell-culture systems, as well as small model organisms.

KEYWORDS:

click chemistry; isotopic labeling; protein engineering; secondary-ion mass spectrometry (SIMS); unnatural amino acid

PMID:
25783034
PMCID:
PMC4471591
DOI:
10.1002/anie.201411692
[Indexed for MEDLINE]
Free PMC Article

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