Department of Medical Biochemistry, Biology and Physics, University of Bari Aldo Moro, Italy.
J Lipid Res. 2010 Sep;51(9):2818-25. doi: 10.1194/jlr.D007328. Epub 2010 Jun 9.
A method of direct lipid analysis by MALDI mass spectrometry in intact membranes, without prior extraction/separation steps, is described. The purple membrane isolated from the extremely halophilic archaeon Halobacterium salinarum was selected as model membrane. Lyophilized purple membrane were grinded with 9-aminoacridine (9-AA) as dry matrix, and the powder mixture was crushed in a mechanical die press to form a thin pellet. Small pieces of the pellet were then attached to the MALDI target and directly analyzed. In parallel, individual archaebacterial phospholipids and glycolipids, together with the total lipid extract of the purple membrane, were analyzed by MALDI-TOF/MS using 9-AA as the matrix in solution. Results show that 9-AA represents a suitable matrix for the conventional MALDI-TOF/MS analysis of lipid extracts from archaeal microorganisms, as well as for fast and reliable direct dry lipid analysis of lyophilized archaebacterial membranes. This method might be of general application, offering the advantage of quickly gaining information about lipid components without disrupting or altering the membrane matrix.
本文描述了一种在完整膜中通过 MALDI 质谱直接进行脂质分析的方法,无需进行预先的提取/分离步骤。选择来自极端嗜盐古菌盐杆菌的紫色膜作为模型膜。将冻干的紫色膜与 9-氨基吖啶(9-AA)一起研磨作为干基质,并将粉末混合物在机械模具压机中粉碎以形成薄的颗粒。然后将颗粒的小块附着在 MALDI 靶上并直接进行分析。同时,使用 9-AA 作为溶液中的基质,通过 MALDI-TOF/MS 对单个古细菌磷脂和糖脂以及紫色膜的总脂质提取物进行分析。结果表明,9-AA 是用于分析古细菌微生物脂质提取物的常规 MALDI-TOF/MS 的合适基质,也是快速可靠地直接分析冻干古细菌膜的干脂质的合适基质。该方法可能具有普遍适用性,具有在不破坏或改变膜基质的情况下快速获取有关脂质成分信息的优势。