Advances in Activity‐Based Sensing Probes for Isoform‐Selective Imaging of Enzymatic Activity

Gardner, Sarah H.; Reinhardt, Christopher J.; Chan, Jefferson

doi:10.1002/anie.202003687

Cited by 57 publications

(40 citation statements)

References 98 publications

(189 reference statements)

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“…31 We envision that activity-based fluorescent sensors can address this gap. [32][33][34][35][36][37] This strategy affords the ability to chemically tune and transform an enzyme's substrate into a fluorescent imaging platform for live-cell applications. To our knowledge, activity-based sensing has not been widely exploited for phenol sulfotransferases in living cells.…”

Section: Main Textmentioning

confidence: 99%

An Activity-Based Fluorescent Sensor for the Detection of the Phenol Sulfotransferase SULT1A1 in Living Cells

Baglia

Mills

Mitra

et al. 2020

Preprint

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The biological activation and incorporation of inorganic sulfate proceeds via a process known as sulfurylation. Transfer of a sulfuryl moiety from the activated sulfate donor, 3’-phosphoadenosine-5’-phosphosulfate (PAPS), to hydroxy-containing substrates by human phenol sulfotransferases (SULT1 family) alters substrate solubility and charge to affect the metabolism of endogenous metabolites, xenobiotics, and drugs. Current methods to monitor SULT1 activity in living cells primarily rely on radiolabeling and/or cell extractions, but these methods do not provide a direct readout of enzyme activity with a dynamic, temporally resolved spatial map in live, intact cells. To fill this gap, here, we present the development, computational modeling, in vitro enzymology, and biological application of Sulfotransferase Sensor-3, STS-3, an activity-based fluorescent sensor for SULT1A1, the most widely expressed and promiscuous SULT1 isoform.

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Section: Main Textmentioning

confidence: 99%

An Activity-Based Fluorescent Sensor for the Detection of the Phenol Sulfotransferase SULT1A1 in Living Cells

Baglia

Mills

Mitra

et al. 2020

Preprint

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“…Recently,Z hang [43,44] and Hong [45] et al embarked on the survey of the physical environment inside aggregated proteins,b ut their work primarily revealed the changes in polarity and viscosity upon protein aggregation using environment-sensitive probes.H owever,i ts till remains elusive how chemically reactive proteins are at their aggregated state. Therefore,c urrent powerful chemical sensing and bioconjugation strategies [46][47][48] may provide hints to address this question.…”

Section: Introductionmentioning

confidence: 99%

“…However, it still remains elusive how chemically reactive proteins are at their aggregated state. Therefore, current powerful chemical sensing and bioconjugation strategies [46–48] may provide hints to address this question.…”

Section: Introductionmentioning

confidence: 99%

Covalent Probes for Aggregated Protein Imaging via Michael Addition

et al. 2021

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Covalent chemical reactions to modify aggregated proteins are rare.Here,wereported covalent Michael addition can generally occur upon protein aggregation. Suchr eactivity was initially discovered by ab ioinspired fluorescent colorswitch probe mimicking the photo-conversion mechanism of Kaede fluorescent protein. This probe was dark with folded proteins but turned on red fluorescence (620 nm) when it noncovalently bound to misfolded proteins.S upported by the biochemical and mass spectrometry results,t he probe chemoselectively reacted with the reactive cysteines of aggregated proteins via covalent Michael addition and gradually switched to green fluorescence (515 nm) upon protein aggregation. Exploiting this Michael addition chemistry in the malachite green dye derivatives demonstrated its general applicability and chemicalt unability,r esulting in different fluorescence color-switch responses.O ur work may offer an ew avenue to explore other chemical reactions upon protein aggregation and design covalent probes for imaging,c hemical proteomics,and therapeutic purposes.

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“…6 Current techniques for directly monitoring NO in vivo have proven useful, but have drawbacks, such as low resolution (EPR), 14,15 poor sensitivity (MRI), 16,17 or invasiveness (amperometry). 18 A complementary approach is the use of activity-based sensing (ABS) probes 19,20 which exploit the chemical reactivity of the target analyte to report on activity with high selectivity. [21][22][23][24] Until recently, most ABS probes for NO have been developed for fluorescence imaging in cellular systems.…”

Section: Introductionmentioning

confidence: 99%

NIR Bioluminescence Probe Enables Discovery of Diet-Induced Modulation of the Tumor Microenvironment via Nitric Oxide

Yadav¹,

Lee²,

Lucero³

et al. 2021

Preprint

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Nitric oxide (NO) plays a critical role in acute and chronic inflammation. NO’s contributions to cancer are of particular interest due to its context-dependent bioactivities. For example, immune cells initially produce cytotoxic quantities of NO in response to the nascent tumor. However, it is believed that this fades over time and reaches a concentration that supports the tumor microenvironment (TME). These complex dynamics are further complicated by other factors, such as diet and oxygenation, making it challenging to establish a complete picture of NO’s impact on tumor progression. Although many activity-based sensing (ABS) probes for NO have been developed, only a small fraction have been employed in vivo and fewer yet are practical in cancer models where the NO concentration is < 200 nM. To overcome this outstanding challenge, we have developed BL660-NO, the first ABS probe for NIR bioluminescence imaging of NO in cancer. Owing to the low intrinsic background, high sensitivity, and deep tissue imaging capabilities of our design, BL660-NO was successfully employed to visualize endogenous NO in cellular systems, a human liver metastasis model, and a murine breast cancer model. Importantly, its exceptional performance facilitated the design of a dietary study to examine the impact of NO on the TME by varying the intake of fat. BL660-NO provides the first direct molecular evidence that intratumoral NO becomes elevated in mice fed a high-fat diet who became obese with larger tumors compared to control animals on a low-fat diet. These results indicate that an inflammatory diet can increase NO production via recruitment of macrophages and overexpression of iNOS which in turn can drive tumor progression.

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Advances in Activity‐Based Sensing Probes for Isoform‐Selective Imaging of Enzymatic Activity

Cited by 57 publications

References 98 publications

An Activity-Based Fluorescent Sensor for the Detection of the Phenol Sulfotransferase SULT1A1 in Living Cells

An Activity-Based Fluorescent Sensor for the Detection of the Phenol Sulfotransferase SULT1A1 in Living Cells

Covalent Probes for Aggregated Protein Imaging via Michael Addition

NIR Bioluminescence Probe Enables Discovery of Diet-Induced Modulation of the Tumor Microenvironment via Nitric Oxide

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