Thioamide quenching of intrinsic protein fluorescence

Goldberg, Jacob M.; Wissner, Rebecca F.; Klein, Alyssa; Petersson, E. James

doi:10.1039/c1cc14708k

Cited by 66 publications

(89 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…33, 34 While intrinisic probes are valuable, they are often limited in their functionality by their need to remain compatible with the translational machinery. Thus, several RS/tRNA pairs derived from the Methanococcus jannaschii ( Mj ) TyrRS/tRNA have been selected for Uaas amenable to post-translational modification through click chemistry reactions, including pXFRS for both azidophenylalanine (Azf, Z) and propargyltyrosine (Ppy, π).…”

Section: Resultsmentioning

confidence: 99%

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

et al. 2016

Self Cite

View full text Add to dashboard Cite

Characterization of the amyloidogenic Parkinson’s Disease protein α-synuclein (αS) has proven difficult due to its structural plasticity. Here, we present a number of complementary methods to site-specifically introduce fluorescent probes to examine αS fibril formation and cellular uptake. By using various combinations of conventional Cys modification, amber codon suppression, transferase mediated N-terminal modification, and native chemical ligation, several variants of singly- and doubly-labeled αS were produced. We validated the nonperturbative nature of the label by a combination of in vitro aggregation kinetics measurements and imaging of the resulting fibrils. The labeled αS can then be used to monitor conformational changes during fibril formation or cellular uptake of αS fibrils in models of disease propagation.

show abstract

Section: Resultsmentioning

confidence: 99%

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…For tryptophan (L-tryptophan, > 98 %, Sigma-Aldrich) on the other hand, pure particles saturated the detector at typical gain settings and produced much higher fluorescence signals than biological materials of comparable size. That the instrument is more sensitive to tryptophan than quinine on a mass basis is likely due to the peak sensitivity of the WIBS PMTs overlapping significantly with the tryptophan emission spectrum and less so with that of quinine (Pant et al, 1990;Goldberg et al, 2012). Therefore, tryptophan-based calibration particles were an internal mixture of tryptophan and ammonium sulfate "filler".…”

Section: Particle Generation and Samplingmentioning

confidence: 99%

Fluorescence calibration method for single-particle aerosol fluorescence instruments

et al. 2017

View full text Add to dashboard Cite

Abstract. Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

show abstract

“…1), which has been probed using many spectroscopic techniques. 2, 6 Here, we investigate the potential for using sensitive hyperpolarized (hp) 129 Xe NMR spectroscopy to detect active CaM in solution.…”

mentioning

confidence: 99%

“…7, 9 The original 26 amino acid sequence was pared down to a 17-mer truncated peptide that retained the ability to bind CaM, which we here call FRRIAR (Scheme 1). 6a This truncated peptide retains two aromatic amino acids characteristic of CaM binding motifs: Phe 1 and Trp 14 in a canonical 1–14 binding mode. 7, 9b …”

mentioning

confidence: 99%

A cryptophane-based “turn-on” ¹²⁹Xe NMR biosensor for monitoring calmodulin

et al. 2017

Self Cite

View full text Add to dashboard Cite

We present the first cryptophane-based “turn-on” 129Xe NMR biosensor, employing a peptide-functionalized cryptophane to monitor the activation of calmodulin (CaM) protein in solution. In the absence of CaM binding, interaction between the peptide and cryptophane completely suppresses the hyperpolarized 129Xe-cryptophane NMR signal. Biosensor binding to Ca2+-activated CaM produces the expected 129Xe-cryptophane NMR signal.

show abstract

Thioamide quenching of intrinsic protein fluorescence

Cited by 66 publications

References 32 publications

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

Fluorescence calibration method for single-particle aerosol fluorescence instruments

A cryptophane-based “turn-on” ¹²⁹Xe NMR biosensor for monitoring calmodulin

Contact Info

Product

Resources

About

Thioamide quenching of intrinsic protein fluorescence

Cited by 66 publications

References 32 publications

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis

Fluorescence calibration method for single-particle aerosol fluorescence instruments

A cryptophane-based “turn-on” 129Xe NMR biosensor for monitoring calmodulin

Contact Info

Product

Resources

About

A cryptophane-based “turn-on” ¹²⁹Xe NMR biosensor for monitoring calmodulin