Joubert F J
Biochim Biophys Acta. 1975 Feb 27;379(2):329-44.
Reduced and S-carboxymethylated phospholipase A (Fraction DE-III) from Naja melanoleuca venom was digested with trypsin, chymotrypsin and thermolysin. The resulting peptides were purified by ion-exchange chromatography on DEAE-cellulose, gel filtration on Sephadex G-25 or G-50 and chromatography and electrophoresis on paper. The amino acid sequences of the intact enzyme and the pur peptides were determined by the Edman procedure, either through the use of the automatic sequencer or by manual manipulation. The chymotryptic digest provided the necessary overlapping peptides which allowed the alignment of the tryptic peptides into a single chain of 119 amino acids. The amino acid sequence of N. melanoleuca phospholipase A shows a high degree of homology with phospholipases A from Bitis gabonica and also from porcine pancreas.
对来自黑曼巴蛇毒液的还原型和S-羧甲基化磷脂酶A（DE-III级分）用胰蛋白酶、胰凝乳蛋白酶和嗜热菌蛋白酶进行消化。所得肽段通过在DEAE-纤维素上进行离子交换色谱、在Sephadex G-25或G-50上进行凝胶过滤以及在纸上进行色谱和电泳进行纯化。完整酶和纯化肽段的氨基酸序列通过埃德曼法测定，可使用自动测序仪或通过手动操作来完成。胰凝乳蛋白酶消化产生了必要的重叠肽段，从而能够将胰蛋白酶肽段排列成一条由119个氨基酸组成的单链。黑曼巴蛇磷脂酶A的氨基酸序列与加蓬咝蝰的磷脂酶A以及猪胰磷脂酶A具有高度同源性。
