Related Experiment Video
Updated: Feb 16, 2026
11:01
Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
Published on: November 23, 2016
10.2K
Activity-Based Probes for Glycosidases: Profiling and Other Applications
Chi-Lin Kuo1, Eline van Meel1, Kassiani Kytidou1
1Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.
Methods in Enzymology
|January 8, 2018
Summary
Researchers developed fluorescent probes to visualize active glucocerebrosidase (GBA) enzyme, aiding Gaucher disease diagnosis and drug discovery. These probes specifically label GBA, enabling sensitive detection in cells and organisms.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Glycosidases are crucial for breaking down glycoconjugates and recycling molecules.
- Lysosomal glycosidase deficiencies cause inherited diseases, notably Gaucher disease, due to impaired glucocerebrosidase (GBA).
- Gaucher disease involves pathological storage of glucosylceramide in macrophages.
Purpose of the Study:
- To develop mechanism-based irreversible inhibitors of GBA with fluorescent reporters.
- To enable specific and sensitive visualization of active GBA molecules.
- To create tools for Gaucher disease diagnosis and drug screening.
Main Methods:
- Utilized cyclophellitol as a lead to design mechanism-based irreversible inhibitors.
- Developed amphiphilic activity-based probes (ABPs) for GBA.
- Employed fluorescent reporters for enzyme visualization.
Main Results:
- Successfully developed fluorescent ABPs for GBA.
- Demonstrated in situ detection of active GBA in cells and organisms.
- Showcased the potential for biochemical diagnosis of Gaucher disease.
Conclusions:
- Fluorescent ABPs are effective tools for visualizing active GBA.
- These probes can aid in Gaucher disease diagnosis and screening for therapeutic compounds.
- The developed concept is extendable to other retaining glycosidases and related diseases.
More Related Videos
Related Concept Videos
01:10Acid–Base Equilibria: Activity-Based Definition of pH
1.3K
For an ideal solution, the pH is defined as the negative logarithm of the hydrogen ion concentration. For a non-ideal solution, an accurate measurement of the pH must consider the negative logarithm of the hydrogen ion activity rather than concentration. In such a solution, the pH can be more accurately defined as the negative logarithm of a product of the hydrogen ion concentration and its activity coefficient.
In solutions of very low ionic strength—for example, pure water—the...
In solutions of very low ionic strength—for example, pure water—the...
1.3K
02:24Ribosome Profiling
4.2K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
4.2K
03:31Labeling DNA Probes
9.5K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
9.5K
02:26tRNA Activation
23.2K
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
23.2K
01:12Taping Over Different Ground Profiles
417
Taping over varying ground profiles requires careful adaptation to achieve accurate measurements. On smooth, level ground with minimal vegetation, the tape can rest directly on the ground. Here, the taping team, typically consisting of a head and a rear tapeman, coordinates their positions with clear communication. The rear tapeman holds the tape at the starting point and guides the head tapeman toward a range pole placed beyond the endpoint, using hand or voice signals to ensure alignment.On...
417
01:26Profile Leveling and Cross Sections
1.7K
Profile leveling and cross-sections are surveying methods used to determine and document terrain elevations for infrastructure projects such as highways, railroads, canals, and pipelines. These methods provide data for earthwork planning and alignment of proposed routes. Profile leveling involves measuring elevations along a fixed line to create a vertical terrain profile. A surveyor sets up a leveling instrument at the benchmark (BM) and records a backsight (BS) to determine the...
1.7K