通过TCEP介导的蛋白质水解:高度选择性的裂变C-终端到酸
Daniel S McCracken1, David J Foreman1, Kyle G Esposito1
1Analytical Research & Development, Merck & Co., Inc., West Point, Pennsylvania, USA.
三二碳乙基 (TCEP) 导致非预期的蛋白质水解在酸残留物,独立于二硫化物键的减少. 这一发现影响了质谱样本准备,并突出了生物化学研究中的潜在工件.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
背景情况:
- 三2-碳氧乙烯) (TCEP) 是一种常见的实验室试剂,用于减少二硫化键.
- 由于其氧化还原潜力,pH范围和处理能力,TCEP比DTT更受欢迎.
- 然而,TCEP的副作用尚未完全理解.
研究的目的:
- 为了调查TCEP的未知副作用.
- 描述TCEP诱导的蛋白质水解的机制.
主要方法:
- 凝电泳是一种凝电泳.
- 质谱测量质量谱测量
主要成果:
- 发现的TCEP会在酸C-末端引起特定位点的蛋白质水解.
- 这种水解发生的独立于二硫化物键的减少.
- 在具有或没有二硫化键或半氨酸的蛋白质中观察到.
结论:
- TCEP可以意外地分裂蛋白质,在质谱学和生物化学研究中创建人工物.
- 这一发现扩大了对TCEP化学的知识,超出了二硫化物还原的范围.
- 为基映射提供了新的可能性,并强调在样本准备过程中要谨慎.
更多相关视频
10:17Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
Published on: January 14, 2020
06:44Hydrolysis of a Ni-Schiff-Base Complex Using Conditions Suitable for Retention of Acid-labile Protecting Groups
Published on: April 6, 2017
相关概念视频
Termination of Translation
Hydrolysis of ATP
If one phosphate group is removed, a molecule of ADP—adenosine diphosphate—remains, along with inorganic phosphate. ADP can be further hydrolyzed to AMP—adenosine...
Acid Halides to Carboxylic Acids: Hydrolysis
As shown below, the mechanism involves a nucleophilic attack by water at the carbonyl carbon to form a tetrahedral intermediate. This is followed by the reformation of the carbon–oxygen π bond along with the departure of a halide ion. A final proton transfer step yields carboxylic...
Amides to Carboxylic Acids: Hydrolysis
Acid-catalyzed hydrolysis:
Hydrolysis of amides under acidic conditions yields carboxylic acids. Since the reaction occurs slowly, hydrolysis requires the conditions of heat.
The mechanism begins with the protonation of the carbonyl oxygen by the acid catalyst. The protonation makes the amide carbonyl carbon more...
Nitriles to Carboxylic Acids: Hydrolysis
Preparation of Carboxylic Acids: Hydrolysis of Nitriles
