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Combining SAMDI Mass Spectrometry and Peptide Arrays to Profile Phosphatase Activities.
- Source :
-
Methods in enzymology [Methods Enzymol] 2018; Vol. 607, pp. 389-403. Date of Electronic Publication: 2018 Jun 19. - Publication Year :
- 2018
-
Abstract
- Phosphatases, the enzymes responsible for dephosphorylating proteins, play critical roles in many cellular processes. While their importance is widely recognized, phosphatase activity and regulation remain poorly understood. Currently, there are few assays available that are capable of directly measuring phosphatase activity and specificity. We have previously introduced SAMDI (self-assembled monolayers on gold for matrix-assisted laser desorption/ionization) mass spectrometry as a technique to profile the substrate specificities of enzymes. SAMDI mass spectrometry assays are well suited to examine phosphatase activities and offer many advantages over current methods. This technique uses monolayers that terminate with a peptide or molecular enzyme substrate and allows for enzyme reactions to be performed on a surface that can easily be rinsed and analyzed by mass spectrometry without the need for analyte labeling. In this chapter, we describe the process of combining SAMDI mass spectrometry with peptide arrays to study the substrate specificities of two protein tyrosine phosphatases.<br /> (© 2018 Elsevier Inc. All rights reserved.)
- Subjects :
- Enzyme Assays instrumentation
Gold chemistry
High-Throughput Screening Assays instrumentation
High-Throughput Screening Assays methods
Peptides analysis
Peptides chemistry
Peptides metabolism
Phosphoric Monoester Hydrolases chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization instrumentation
Substrate Specificity
Enzyme Assays methods
Phosphoric Monoester Hydrolases metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
Subjects
Details
- Language :
- English
- ISSN :
- 1557-7988
- Volume :
- 607
- Database :
- MEDLINE
- Journal :
- Methods in enzymology
- Publication Type :
- Academic Journal
- Accession number :
- 30149867
- Full Text :
- https://doi.org/10.1016/bs.mie.2018.04.021