Separation of phospholipase A2 in Habu snake venom by glycyrrhizin (GL)-affinity column chromatography and identification of a GL-sensitive enzyme
Separation of phospholipase A2 in Habu snake venom by glycyrrhizin (GL)-affinity column chromatography and identification of a GL-sensitive enzyme
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DOI：
10.1248/bpb.21.574
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发表时间：
1998-06-01
影响因子：
2
通讯作者：
Takasaki, C
中科院分区：
文献类型：
--
作者：
Ohtsuki, K;Abe, Y;Takasaki, C
通过甘草甜素（GL）亲和层析和Mono S柱色谱（HPLC），选择性纯化了哈布蛇毒的两个Superdex级分（P-II和P-III级分）中至少四种GL结合蛋白（p25、p17、p15-1和p15-2），Bg测定了它们的N端部分氨基酸序列， 鉴定出金属蛋白酶 (p25) 和三种 GL 结合磷脂酶 Az (gbPLA(2)s) [PA2Y (p17)、PA21 (p15-1) 和 PA2B (p15-2)]。 PA2B（赖氨酸-49 PLA(2)）被发现对 GL 最敏感，因为 (i) 它与 GL 亲和柱强烈结合； (ii)其酶活性被低剂量(ID50＝约1.5μM)的GL选择性抑制，但不被GA抑制。此外，这三种gbPLA(2)在体外被酪蛋白激酶II (CK-II)磷酸化，GL在体外抑制CK-II介导的酶活性刺激。
By means of glycyrrhizin (GL)-affinity and Mono S column chromatographies (HPLC), at least four GL-binding proteins (p25, p17, p15-1 and p15-2) in the two Superdex fractions (P-II and P-III fractions) from Habu snake venom were selectively purified, Bg determination of their N-terminal partial amino acid sequences, a metalloprotease (p25) and three GL-binding phospholipases Az (gbPLA(2)s) [PA2Y (p17), PA21 (p15-1) and PA2B (p15-2)] were identified. PA2B (lysine-49 PLA(2)) was found to be the most sensitive to GL because (i) it strongly bound to a GL-affinity column; and (ii) its enzyme activity was selectively inhibited by low dose (ID50=approx. 1.5 mu M) of GL, hut not by GA. Furthermore, these three gbPLA(2)s were phosphorylated by casein kinase II (CK-II) in vitro and GL inhibited the CK-II-mediated stimulation of their enzyme activities in vitro.