Activity based proteomics

Activity based proteomics

Activity based proteomics, or activity based protein profiling (ABPP) is a functional proteomic technology that uses specially designed chemical probes that react with mechanistically-related classes of enzymes [Berger AB, et al. "Activity-based protein profiling: applications to biomarker discovery, in vivo imaging and drug discovery." American Journal of Pharmacogenomics 2004 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=search&DB=pubmed Article] ] . The basic unit of ABPP is the probe which typically consists of two elements: a reactive group (RG) and a tag. Additionally, some probes may contain a binding group which enhances selectivity. The reactive group usually contains an electrophile that gets covalently-linked to a nucleophilic residue in the active site of an active enzyme. An enzyme that is inhibited by enzyme inhibitors or post-translational modifications will not react with an activity-based probe. The tag may be either a reporter such as a fluorophore or a "handle" such as biotin or an alkyne or azide for use with the Huisgen 1,3-dipolar cycloaddition (also known as click chemistry) [Speers AE, et al. "Activity-Based Protein Profiling in Vivo Using a Copper(I)-Cataqlyzed Azide-Alkyne [3+2] Cycloaddition" Journal of the American Chemical Society 2003 [http://www.scripps.edu/cb/cravatt/pdf/Speersetal2003.pdf Article] ] .

A major advantage of ABPP is the ability to monitor enzyme "activity" directly, rather than being limited to protein or mRNA abundance. With classes of enzymes such as the serine proteases [ Liu Y, et al. "Activity-based protein profiling: The serine hydrolases" Proceedings of the National Academy of Sciences 1999 [http://www.pnas.org/cgi/content/abstract/96/26/14694 Article] ] and metalloproteases [ Saghatelian A , et al. "Activity-based probes for the proteomic profiling of metalloproteases" Proceedings of the National Academy of Sciences 2004 [http://www.pnas.org/cgi/content/abstract/101/27/10000 Article] ] that often interact with endogenous inhibitors or that exist as inactive zymogens, this technique offers a valuable advantage over traditional techniques that rely on abundance rather than activity.

Finally, in recent years ABPP has been combined with tandem mass spectrometry enabling the identification of hundreds of active enzymes from a single sample. This technique, known as "ABPP-MudPIT" is especially useful for profiling inhibitor selectivity as the potency of an inhibitor can be tested against hundreds of targets simultaneously.

ABPP was developed by in the late 1990s and early 2000s by Benjamin Cravatt at The Scripps Research Institute, [http://bogyolab.stanford.edu/ Matthew Bogyo] at Stanford, and others.

ee also

* Mass spectrometry
* Proteomics
* Related inhibitors MAFP and DIFP

References


Wikimedia Foundation. 2010.

Игры ⚽ Нужна курсовая?

Look at other dictionaries:

  • Chemical biology — is a scientific discipline spanning the fields of chemistry and biology that involves the application of chemical techniques and tools, often compounds produced through synthetic chemistry, to the study and manipulation of biological systems.… …   Wikipedia

  • Enzyme inhibitor — Enzyme inhibitors are molecules that bind to enzymes and decrease their activity. Since blocking an enzyme s activity can kill a pathogen or correct a metabolic imbalance, many drugs are enzyme inhibitors. They are also used as herbicides and… …   Wikipedia

  • Benjamin Cravatt III — Benjamin F Cravatt Fields Chemical Biology Institutions The Scripps Research Institute ActivX Biosciences …   Wikipedia

  • Organophosphorus — compounds are degradable organic compounds containing carbon–phosphorus bonds (thus excluding from phosphate and phosphite esters, which lack such kind of bonding), used primarily in pest control as an alternative to chlorinated hydrocarbons that …   Wikipedia

  • Active site — The active site of an enzyme contains the catalytic and binding sites. The structure and chemical properties of the active site allow the recognition and binding of the substrate.The active site is usually a small pocket at the surface of the… …   Wikipedia

  • Serine hydrolase — The serine hydrolase superfamily is one of the largest known enzyme families comprising approximately 1% of the genes in the human genome. This family includes:* serine proteases like trypsin * lipases like pancreatic lipase, hormone sensitive… …   Wikipedia

  • Minimum Information Standards — Minimum Information (MI) standards or reporting guidelines specify the minimum amount of meta data (information) and data required to meet a specific aim or aims. Usually the aim is to provide enough meta data and data to enable the unambiguous… …   Wikipedia

  • Phosphorylation — is the addition of a phosphate (PO4) group to a protein molecule or a small molecule. It can also be thought of as the introduction of a phosphate group into an organic molecule . Its prominent role in biochemistry is the subject of a very large… …   Wikipedia

  • Blood proteins — Blood proteins, also called serum proteins , are proteins found in blood plasma. Serum total protein in blood is 7g/dl, which in total makes 7% of total body weight. They serve many different functions, including *circulatory transport molecules… …   Wikipedia

  • Natural scientific research in Canada — This article outlines the history of natural scientific research in Canada, including mathematics, physics, astronomy, space science, geology, oceanography, chemistry, biology, medical research and psychology. The social sciences are not treated… …   Wikipedia

Share the article and excerpts

Direct link
Do a right-click on the link above
and select “Copy Link”