Chapter title |
Applying Arginylation for Bottom-Up Proteomics.
|
---|---|
Chapter number | 16 |
Book title |
Protein Arginylation
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2935-1_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2934-4, 978-1-4939-2935-1
|
Authors |
H. Alexander Ebhardt |
Editors |
Anna S. Kashina |
Abstract |
Arginylation is an enzymatic reaction in which arginyl-tRNA protein transferase 1 (ATE1, EC 2.3.2.8) conjugates a single arginyl moiety from aminoacylated tRNA(Arg) onto a target polypeptide. We established arginylation for in vitro labeling of peptides with N-terminal acidic amino acids. Consistent with prior knowledge, arginylated peptides flanked by basic amino acids result in rich redundant MS/MS fragment spectra using various precursor fragmentation modes. Arginylation carried out by ATE1 is a fast method for labeling peptides. Sequence-specific proteolytic digest of proteins is best carried out using a double digest of proteins by Lys-C and Asp-N to generate peptides with a basic amino acid on the C-terminus and an acidic amino acid on the N-terminus. Under these conditions, arginylation is specific for N-terminal acidic amino acids and results in a near 2× sequence coverage in the MS/MS spectrum are achieved. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 3 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 1 | 33% |
Lecturer | 1 | 33% |
Student > Postgraduate | 1 | 33% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 67% |
Agricultural and Biological Sciences | 1 | 33% |