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Can I read Analysis of Isoaspartate in Peptides and Proteins without the Use of Radioisotopes on EtoBox?
Analysis of Isoaspartate in Peptides and Proteins without the Use of Radioisotopes by Brandon T. Schurter; Dana W. Aswad is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Analysis of Isoaspartate in Peptides and Proteins without the Use of Radioisotopes about?
A rapid and sensitive HPLC-based method for quantitating isoaspartate levels in peptides and proteins is described. The analyte is incubated for 40 min with S-adenosyl-L-methionine and the commercially available enzyme protein L-isoaspartyl methyltransferase. Methylation of isoaspartyl sites results in stoichiometric production of S-adenosyl-L-homocysteine that is separated from the other components of the reaction by reversed-phase HPLC and quantitated online by absorbance at 260 nm. This method can accurately detect 5 pmol or less of isoaspartate and works with tryptic digests as well as intact proteins. Using a commercially available isoaspartyl peptide, the relationship between isoaspartate levels and S-adenosyl-L-homocysteine production was found to be linear and stoichiometric over a range of 5-250 pmol. Compared to methods that measure [ 3 H]methanol production after methylation with S-adenosyl-L-[methyl-3 H]methionine, the HPLC method is safer, faster, less expensive, and equally sensitive.
Who reads Analysis of Isoaspartate in Peptides and Proteins without the Use of Radioisotopes?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Brandon T. Schurter; Dana W. Aswad
- Publisher
- Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)
- Published
- 2000
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)