|
|||||
|
|
A tris (2-carboxyethyl) phosphine (TCEP) related cleavage on cysteine-containing proteins |
|
| Authors: | Peiran Liu Brian W O’Mara Bethanne M Warrack Wei Wu Yunping Huang Yihong Zhang Rulin Zhao Mei Lin Michael S Ackerman Peter K Hocknell Guodong Chen Li Tao Siegfried Rieble Jack Wang David B Wang-Iverson Adrienne A Tymiak Michael J Grace Reb J Russell |
| Institution: | 1. Department of Biologics Product and Process Development, Bristol-Myers Squibb Technical Operations, Princeton, New Jersey, USA 2. Department of Bioanalytical and Discovery Analytical Sciences, Bristol-Myers Squibb Research and Development, Princeton, New Jersey, USA 3. Adnexus, a Bristol-Myers Squibb Research and Development company, Waltham, Massachusetts, USA 4. Department of Chemical Synthesis, Bristol-Myers Squibb Research and Development, Princeton, New Jersey, USA |
| Abstract: | Introduced in the late 1980s as a reducing reagent, Tris (2-carboxyethyl) phosphine (TCEP) has now become one of the most widely used protein reductants. To date, only a few studies on its side reactions have been published. We report the observation of a side reaction that cleaves protein backbones under mild conditions by fracturing the cysteine residues, thus generating heterogeneous peptides containing different moieties from the fractured cysteine. The peptide products were analyzed by high performance liquid chromatography and tandem mass spectrometry (LC/MS/MS). Peptides with a primary amine and a carboxylic acid as termini were observed, and others were found to contain amidated or formamidated carboxy termini, or formylated or glyoxylic amino termini. Formamidation of the carboxy terminus and the formation of glyoxylic amino terminus were unexpected reactions since both involve breaking of carbon—carbon bonds in cysteine. |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ | |