Joubert F J
Biochim Biophys Acta. 1975 Feb 27;379(2):345-59.
The complete amino acid sequences of phospholipase A (Fractions DE-I and DE-II) from Naja melanoleuca (Forest cobra) have been elucidated. The reduced and S-carboxymethylated isoenzyme were digested with trypsin and thermolysin and the peptides were purified by ion-exchange chromatography, gel filtration and chromatography or electrophoresis on paper. The Edman procedure, either through the use of the automatic sequencer or by manual manipulation, was employed to obtain the sequences of the intact isoenzymes and the pure peptidesmthe thermolysin digest provided the necessary overlapping peptides which allowed the alignment of the tryptic peptides of Fraction DE-I. The tryptic peptides of Fraction DE-II were either identical or homologous to the tryptic peptides of Fraction I and Fraction III [12] and were aligned in the same order as that of Fractions DE-I or DE-III. The amino acid sequence of N. melanoleuca phospholipase A, Fraction I, shows a high degree of homology with Fraction DE-II and also with Fraction DE-III, previously reported on [12].
黑曼巴蛇（森林眼镜蛇）磷脂酶A（DE-I和DE-II组分）的完整氨基酸序列已被阐明。将还原型和S-羧甲基化的同工酶用胰蛋白酶和嗜热菌蛋白酶消化，肽段通过离子交换色谱、凝胶过滤以及纸层析或电泳进行纯化。采用埃德曼降解法，通过自动测序仪或手动操作来获得完整同工酶和纯肽段的序列；嗜热菌蛋白酶消化产物提供了必要的重叠肽段，从而能够对DE-I组分的胰蛋白酶肽段进行比对。DE-II组分的胰蛋白酶肽段与I组分和III组分[12]的胰蛋白酶肽段相同或同源，并按照与DE-I或DE-III组分相同的顺序排列。黑曼巴蛇磷脂酶A的I组分氨基酸序列与DE-II组分以及先前报道的[12]DE-III组分具有高度同源性。
