Lu R C, Elzinga M
Biochemistry. 1977 Dec 27;16(26):5801-6. doi: 10.1021/bi00645a025.
Actin was purified from calf brains by chromatography on DEAE-Sephadex and hydroxylapatite. The protein was then subjected to amino acid sequence analysis by isolating and sequencing its cyanogen bromide peptides. CB-1, 3, 4, 5, 6, 9, 10, and 12 correspond to equivalent segments of rabbit skeletal muscle actin, while subsitutions involving methionines give rise to some new peptides. The region that corresponds to CB-13 in muscle actin becomes two peptides in the brain protein because of a Leu leads to Met replacement at position 16, while Met leads to Leu substitutions at positions 176 and 298 give rise to two larger peptides, CB-15 + 7 and CB-8 + 2, which correspond to muscle actin CB-15 fused with CB-7 and CB-8 fused with CB-2, respectively. The peptides that have been isolated from brain actin contain 267 of the 374 residues in actin, of which 157 have been unequivocally identified. When the data are compared with those for rabbit skeletal muscle actin, 11 replacements are seen; thus the two actins differ at about 7% of the positions examined.
通过在DEAE - 葡聚糖凝胶和羟基磷灰石上进行色谱法从牛脑中纯化肌动蛋白。然后通过分离和测序其溴化氰肽段对该蛋白质进行氨基酸序列分析。CB - 1、3、4、5、6、9、10和12对应于兔骨骼肌肌动蛋白的等效片段,而涉及甲硫氨酸的替换产生了一些新的肽段。由于在第16位亮氨酸被甲硫氨酸取代,在肌肉肌动蛋白中对应于CB - 13的区域在脑蛋白中变成了两个肽段,而在第176位和第298位甲硫氨酸被亮氨酸取代产生了两个更大的肽段,CB - 15 + 7和CB - 8 + 2,它们分别对应于与CB - 7融合的肌肉肌动蛋白CB - 15和与CB - 2融合的CB - 8。从脑肌动蛋白中分离出的肽段包含肌动蛋白374个残基中的267个,其中157个已被明确鉴定。当将这些数据与兔骨骼肌肌动蛋白的数据进行比较时,发现了11个替换;因此,这两种肌动蛋白在所检查位置的约7%处存在差异。