低分子量牛心蛋白脂质的纯化与特性分析：运用快原子轰击质谱法进行鉴定

Purification and characterization of low-molecular-weight beef heart proteolipids: use of fast atom bombardment-mass spectrometry for identification.

作者信息

Boyot P, Trifilieff E, Van Dorsselaer A, Luu B

机构信息

Laboratoire de Chimie Organique des Substances Naturelles, UA CNRS 31, Strasbourg, France.

出版信息

Anal Biochem. 1988 Aug 15;173(1):75-85. doi: 10.1016/0003-2697(88)90162-5.

DOI:10.1016/0003-2697(88)90162-5

PMID:2903696

Abstract

Bovine cardiac muscle was extracted by an acidic chloroform/methanol mixture. A combination of gel permeation and ion-exchange chromatographies in organic solvents and HPLC allowed the purification of subunits VIIIa (Mr 5400) and VIIIb (Mr 4900) of cytochrome c oxidase and of A6L protein (Mr 7900) of ATP synthase. The identification of the proteins was made possible by measurement of their molecular weight by fast atom bombardment-mass spectrometry (FAB-MS) in conjunction with conventional Edman degradation. The determination by FAB-MS of the molecular weight of A6L protein confirmed its supposed formylated N-terminal methionine.

摘要

用酸性氯仿/甲醇混合物提取牛心肌。在有机溶剂中结合凝胶渗透色谱法和离子交换色谱法以及高效液相色谱法，可纯化细胞色素c氧化酶的VIIIa亚基（分子量5400）和VIIIb亚基（分子量4900）以及ATP合酶的A6L蛋白（分子量7900）。通过快速原子轰击质谱法（FAB-MS）结合传统的埃德曼降解法测量其分子量，从而实现对这些蛋白质的鉴定。通过FAB-MS测定A6L蛋白的分子量，证实了其假定的N端甲酰化甲硫氨酸。