Eremin V A, Chekulaeva L N, Kharat'ian E F, Tabagua I I, Ostrovskiĭ D N
Mikrobiologiia. 1978 Nov-Dec;47(6):1037-43.
A technique has been elaborated for preparation of deuterated membranes from deuterated cells of Micrococcus lysodeikticus containing 85--90% of deuterium according to the data of IR and PMR spectroscopy. Normal lysis of the deuterated cells of M. lysodeikticus requires a concentration of lysozyme which is eight times higher than for usual cells (8 mg per 1 g of wet deuterated cells) and an addition of the lytic enzymes E-2 (2 mg per 1 g of wet deuterated cells). Preparations of deuterated membranes purified from ribosomes and proteins can be obtained by treating of the lysate with RNAase and washing in 0.5 M NaCl. The purity of the deuterated membranes was evaluated by the evidence of electron microscopy, IR spectra and enzyme activities. After hydrogen atoms were substituted by deuterium, the secondary structure of the total membrane protein, the ratio between the activities of the respiratory chain enzymes, and the relative content of the lipid and protein components of the deuterated membranes remain at the same level as in the protonated ones.
已研发出一种技术,可根据红外光谱和核磁共振光谱数据,从含85%-90%氘的溶壁微球菌的氘代细胞制备氘代膜。溶壁微球菌的氘代细胞正常裂解所需的溶菌酶浓度比普通细胞高八倍(每1克湿氘代细胞8毫克),并需添加溶菌酶E-2(每1克湿氘代细胞2毫克)。通过用核糖核酸酶处理裂解物并在0.5M氯化钠中洗涤,可获得从核糖体和蛋白质中纯化的氘代膜制剂。通过电子显微镜、红外光谱和酶活性证据评估氘代膜的纯度。氢原子被氘取代后,氘代膜总膜蛋白的二级结构、呼吸链酶活性之间的比率以及脂质和蛋白质成分的相对含量与质子化膜中的水平保持相同。