関連する実験動画

Updated: Feb 15, 2026

Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms

Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms

Published on: January 25, 2018

TCEP媒介タンパク質水解:高度に選択的な割れ目C-ターミナルからアスパルティック酸まで

Daniel S McCracken¹, David J Foreman¹, Kyle G Esposito¹

¹Analytical Research & Development, Merck & Co., Inc., West Point, Pennsylvania, USA.

Rapid communications in mass spectrometry : RCM

|February 14, 2026

まとめ

Tris(2-カルボキシエチル) フォスフィン (TCEP) は,ディスルフィード結合の減少とは独立して,アスパルチン酸残留で予期せぬタンパク質水解を引き起こします. この発見は,質量スペクトロメトリーサンプル準備に影響を与え,生化学研究における潜在的な人工物を強調しています.

科学分野:

バイオケミストリーバイオケミストリー
アナリティカル・ケミストリー (Analytical Chemistry) とは

背景:

Tris ((2-カルボキシエチル) フォスフィン (TCEP) は,二硫化結合を減少させるための一般的な実験室用反応剤です.
TCEPは,その酸化還元能力,pHの範囲,および処理により,DTTよりも好ましい.
しかし,TCEPの副作用は完全に理解されていません.

研究の目的:

TCEPの未知の副作用を調査する.
TCEP誘発タンパク質水解のメカニズムを特徴づけるために.

主な方法:

ゲル電泳によるゲル電泳です.
質量スペクトロメトリによる質量スペクトロメトリです.

主要な成果:

発見されたTCEPは,アスパルティック酸C-末端でサイト固有のタンパク質の水解を引き起こします.
この水解は,二硫化物結合の減少とは無関係に発生する.
ディスルファイド結合またはシステインを含有または含有しないタンパク質で観察される.

結論:

TCEPは予期せぬ形でタンパク質を分裂させ,質量スペクトロメトリや生化学研究における人工物を作り出すことができる.
この発見は,二酸化硫化物の還元を超えて,TCEPの化学に関する知識を広げています.
ペプチドマッピングの新たな可能性を提供し,サンプル準備における注意を強調する.

さらに関連する動画

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

Hydrolysis of a Ni-Schiff-Base Complex Using Conditions Suitable for Retention of Acid-labile Protecting Groups

Hydrolysis of a Ni-Schiff-Base Complex Using Conditions Suitable for Retention of Acid-labile Protecting Groups

Published on: April 6, 2017

関連する概念動画

Termination of Translation

Termination of Translation

The large ribosomal subunit has several important structures essential to translation. These include the peptidyl transferase center (PTC) - which is the site where the peptide bond is formed - and a large, internal, water-filled tube through which the nascent polypeptide moves. This latter structure is called the Peptide Exit Tunnel, and it begins at the PTC and spans the body of the large ribosomal subunit. During translation, as the nascent polypeptide chain is synthesized, it passes through...

Hydrolysis of ATP

Hydrolysis of ATP

The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
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

Acid Halides to Carboxylic Acids: Hydrolysis

Hydrolysis of acid halides is a nucleophilic acyl substitution reaction in which acid halides react with water to give carboxylic acids. The reaction occurs readily and does not require acid or a base catalyst.
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

Amides to Carboxylic Acids: Hydrolysis

Amides can undergo either acid-catalyzed hydrolysis or base-promoted hydrolysis through a typical nucleophilic acyl substitution. Each hydrolysis requires severe conditions.
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

Nitriles to Carboxylic Acids: Hydrolysis

Nitriles undergo acid-catalyzed hydrolysis or base-catalyzed hydrolysis to form a carboxylic acid. These reactions proceed via an amide intermediate.

Preparation of Carboxylic Acids: Hydrolysis of Nitriles

Preparation of Carboxylic Acids: Hydrolysis of Nitriles

Nitriles (R–CN) can be converted into carboxylic acids (R–COOH) upon treatment with aqueous acids, i.e., upon hydrolysis of nitriles. Under base-catalyzed conditions, carboxylate anions (R–COO−) are formed.