Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette cedex, France.
Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette cedex, France.
Methods Enzymol. 2023;684:135-166. doi: 10.1016/bs.mie.2023.02.016. Epub 2023 Mar 28.
N-terminal myristoylation is an essential eukaryotic modification crucial for cellular homeostasis in the context of many physiological processes. Myristoylation is a lipid modification resulting in a C14 saturated fatty acid addition. This modification is challenging to capture due to its hydrophobicity, low abundance of target substrates, and the recent discovery of unexpected NMT reactivity including myristoylation of lysine side chains and N-acetylation in addition to classical N-terminal Gly-myristoylation. This chapter details the high-end approaches developed to characterize the different features of N-myristoylation and its targets through in vitro and in vivo labeling.
N-端豆蔻酰化是一种必需的真核修饰,对于许多生理过程中的细胞内稳态至关重要。豆蔻酰化是一种脂质修饰,导致添加一个 C14 饱和脂肪酸。由于其疏水性、靶底物的低丰度以及最近发现的出乎意料的 NMT 反应性,包括赖氨酸侧链的豆蔻酰化和除经典的 N-端甘氨酸豆蔻酰化之外的 N-乙酰化,这种修饰很难被捕捉到。本章详细介绍了通过体外和体内标记来描述 N-豆蔻酰化及其靶标的不同特征的高端方法。