Garrett Teresa A, Kordestani Reza, Raetz Christian R H
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, USA.
Methods Enzymol. 2007;433:213-30. doi: 10.1016/S0076-6879(07)33012-7.
Cardiolipin (CL), a tetra-acylated glycerophospholipid composed of two phosphatidyl moieties linked by a bridging glycerol, plays an important role in mitochondrial function in eukaryotic cells. Alterations to the content and acylation state of CL cause mitochondrial dysfunction and may be associated with pathologies such as ischemia, hypothyrodism, aging, and heart failure. The structure of CL is very complex because of microheterogeneity among its four acyl chains. Here we have developed a method for the quantification of CL molecular species by liquid chromatography-electrospray ionization mass spectrometry. We quantify the M-2H ion of a CL of a given molecular formula and identify the CLs by their total number of carbons and unsaturations in the acyl chains. This method, developed using mouse macrophage RAW 264.7 tumor cells, is broadly applicable to other cell lines, tissues, bacteria and yeast. Furthermore, this method could be used for the quantification of lyso-CLs and bis-lyso-CLs.
心磷脂（CL）是一种由两个通过桥连甘油连接的磷脂酰部分组成的四酰化甘油磷脂，在真核细胞的线粒体功能中起着重要作用。CL含量和酰化状态的改变会导致线粒体功能障碍，并可能与缺血、甲状腺功能减退、衰老和心力衰竭等病理状况相关。由于其四条酰基链之间存在微观异质性，CL的结构非常复杂。在此，我们开发了一种通过液相色谱 - 电喷雾电离质谱法定量CL分子种类的方法。我们对给定分子式的CL的M - 2H离子进行定量，并通过其酰基链中的总碳原子数和不饱和度来鉴定CL。该方法是使用小鼠巨噬细胞RAW 264.7肿瘤细胞开发的，广泛适用于其他细胞系、组织、细菌和酵母。此外，该方法可用于定量溶血心磷脂和双溶血心磷脂。
