CAJ | 학술논문

通过对纯化得到的长白山白眉蝮蛇蛇毒磷脂酶A_2(ABUSV-PLA_2)进行胶内胰蛋白酶酶解,产生的肽段经高效液相色谱-电喷雾串联质谱(HPLC-nESI-MS/MS)进行序列测定,蛋白鉴定采用Sequest Bioworks软件完成.ABUSV-PLA_2与其它蛇毒来源PLA_2的氨基酸序列比对表明:ABUSV-PLA_2是一种新的蛇毒酸性磷脂酶A_2.以ADP诱导的人血小板聚集实验结果表明:ABUSV-PLA_2对ADP诱导的血小板聚集有轻微的解凝效应,但具有显著拮抗血小板的聚集作用,并呈现明显的剂量-效应关系,IC50为356 nmol/L.
통과대순화득도적장백산백미복사사독린지매A_2(ABUSV-PLA_2)진행효내이단백매매해,산생적태단경고효액상색보-전분무천련질보(HPLC-nESI-MS/MS)진행서렬측정,단백감정채용Sequest Bioworks연건완성.ABUSV-PLA_2여기타사독래원PLA_2적안기산서렬비대표명:ABUSV-PLA_2시일충신적사독산성린지매A_2.이ADP유도적인혈소판취집실험결과표명:ABUSV-PLA_2대ADP유도적혈소판취집유경미적해응효응,단구유현저길항혈소판적취집작용,병정현명현적제량-효응관계,IC50위356 nmol/L.
An acidic phospholipase A_2 obtained form Chinese Agkistrodon blomhoffii ussurensis snake venom,denoted as ABUSVPLA_2,was digested by trypsin in-gel.The tryptic peptides were separated by using high performance liquid chromatography (HPLC) through a C18 column and sequenced consequently by the electro-spray ionization tandem mass spectrometry (ESI-MS/MS).Protein identification was performed by searching the mass spectrometry data against the NCBInr database through the Bioworks software.Sequence comparison between the ABUSV-PLA_2 and other snake venoms sourced phospholipase A_2s indicated that ABUSV-PLA_2 is a novel snake venom phospholipase A_2.Results from human platelet aggregation experiments indicated that ABSUV-PLA_2 slightly affects the dissociation of aggregated platelet induced by ADP,however,ABUSV-PLA_2 strongly inhibits the human platelet aggregation and its inhibition is dose-dependent.The IC50 is 356 nmol/L.