Molecular characterization and anticoagulant activity of a novel annexin derived from the Taenia solium.
A novel annexin gene was isolated from the Taenia solium cDNA library by degenerate PCR. The purified protein was generated by hydrolysis with thrombin in glutathione S-transferase (GST) affinity chromatography following expression in GST tagged pGEX-5T vector. It shares the common structural features of the annexin family and specially possesses two unique fragments between repeating domains II and III which do not exist in other annexin family members revealed by aligning and homology modeling, hence it was designated as Tso ANXB2 according to the new nomenclature of annexins. According to the parasitic behaviors of the origin, a series of coagulation assays was performed, indicating that Tso ANXB2 inhibits extrinsic blood coagulation pathway and platelet aggregation also has platelet binding activity. Nevertheless, the mutant protein deleting the consensus coagulation-related KGD motif of Tso ANXB2 showed significant decreasing platelet binding and anticoagulation activity.
Wang
KaiHui
K
Department of Medical Genetics, The Second Military Medical University, Xiang Yin Road 800, Shanghai 200433, PR China.
Guo
YingJun
Y
Li
Ka
K
Lu
YiMing
Y
Zhang
Yi
Y
Sun
ShuHan
S
Yan
HongLi
H
Zhang
Shu
S
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D000925:Anticoagulants / Q000494:pharmacology*; D001483:Base Sequence; D001777:Blood Coagulation / Q000187:drug effects / Q000502:physiology*; D003001:Cloning, Molecular; D015151:Immunoblotting; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010314:Partial Thromboplastin Time; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology; D011487:Protein Conformation; D011517:Prothrombin Time; D018510:RNA, Helminth / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D041201:Taenia solium / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D017302:Annexins; D000925:Anticoagulants; D018510:RNA, Helminth; D011994:Recombinant Proteins