Functional characterization of a salivary apyrase from the sand fly, Phlebotomus duboscqi, a vector of Leishmania major
Functional characterization of a salivary apyrase from the sand fly, Phlebotomus duboscqi, a vector of Leishmania major
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DOI：
10.1016/j.jinsphys.2009.07.010
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发表时间：
2009-11-01
影响因子：
2.2
通讯作者：
Valenzuela, Jesus G.
中科院分区：
文献类型：
--
作者：
Hamasaki, Ryoichi;Kato, Hirotomo;Valenzuela, Jesus G.
通过对一种白头翁(Phlebotomus(P.)杜波斯齐唾液腺cdna文库。序列分析表明，这些分子中与酶活性有关的氨基酸，包括核苷酸酶活性和钙与核苷酸的结合，都是非常保守的。在大肠杆菌中表达并纯化了重组杜氏杆菌唾液apyrase。该蛋白能有效地降解ADP和ATP，但不能以钙依赖的方式降解AMP、GDP、CDP或UDP。此外，重组蛋白还能抑制ADP和胶原诱导的血小板聚集。结果表明，该唾液蛋白在杜氏狸殖吸虫的摄血过程中起着重要作用。其独特的酶活性使唾液apyrase成为治疗血栓病理的有吸引力的候选药物，以及用于广泛研究目的的试剂。(C)2009爱思唯尔有限公司。保留所有权利。
Two transcripts coding for proteins homologous to apyrases were identified by massive sequencing of a Phlebotomus (P.) duboscqi salivary gland cDNA library. The sequence analysis revealed that the amino acids important for enzymatic activity including nucleotidase activity and the binding of calcium and nucleotides were well conserved in these molecules. A recombinant P. duboscqi salivary apyrase was expressed in Escherichia coli and purified. The resulting protein efficiently hydrolyzed ADP and ATP, but not AMP, GDP, CDP or UDP, in a calcium-dependent manner. Further, the recombinant protein inhibited ADP- and collagen-induced platelet aggregation. The results indicated that this salivary protein plays an important role in the blood-feeding process in P. duboscqi. Its unique enzymatic activity makes the salivary apyrase an attractive candidate as a therapeutic agent for the treatment of thrombotic pathologies as well as a reagent for a wide variety of research purposes. (C) 2009 Elsevier Ltd. All rights reserved.