| Chapter title |
[14C]-Acetyl-Coenzyme A-Based In Vitro N-Terminal Acetylation Assay
|
|---|---|
| Chapter number | 1 |
| Book title |
Protein Terminal Profiling
|
| Published in |
Methods in molecular biology, March 2017
|
| DOI | 10.1007/978-1-4939-6850-3_1 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6849-7, 978-1-4939-6850-3
|
| Authors |
Adrian Drazic, Thomas Arnesen |
| Editors |
Oliver Schilling |
| Abstract |
N-terminal acetylation is one of the most abundant co- and posttranslational protein modifications, conserved from prokaryotes to eukaryotes. The functional consequences of this modification are manifold, ranging from protein folding, stability, and interaction to subcellular localization. We describe here an isotope-labeled [(14)C]-acetyl-Coenzyme A-based acetylation assay, allowing the determination of weak catalytic activities of NATs in vitro. It allows the use of purified recombinant enzymes from Escherichia coli, or co-immunoprecipitated enzymes from various organisms, as well as the determination of the in vitro activity of various cell lysates. Although marked as an old-fashioned biochemical approach, it is the ideal method to hunt for catalytic activities and defining peptide specificities of new potential N-terminal acetyltransferase candidates. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 4 | 27% |
| Student > Ph. D. Student | 2 | 13% |
| Student > Bachelor | 2 | 13% |
| Unspecified | 1 | 7% |
| Student > Doctoral Student | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 4 | 27% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 5 | 33% |
| Agricultural and Biological Sciences | 2 | 13% |
| Chemistry | 2 | 13% |
| Immunology and Microbiology | 1 | 7% |
| Unspecified | 1 | 7% |
| Other | 0 | 0% |
| Unknown | 4 | 27% |