Molecular cloning and sequence analysis of alboaggregin B.
Green pit viper (Trimeresurus albolabris) venom contains a variety of C-type lectin-like proteins (CLPs) causing platelet aggregation and consumptive thrombocytopenia in biting victims. Alboaggregin B (AL-B), a heterodimeric glycoprotein (Gp) Ib-binding protein, was purified from the venom, but there is no reported cDNA sequence and the platelet agglutination mechanism is poorly understood. The full-length AL-B beta clone was obtained from T. albolabris venom gland cDNA library. AL-B alpha was, later, derived using 3'-RACE based on the conserved sequence. In this study, purified AL-B dimer agglutinated human platelets with the EC(50) of 180 nM and was completely inhibited by anti GpIb antibody. MALDI ToF mass spectroscopy found no glycosylation. The peptide mass fingerprints were matched with deduced amino acid sequences of cloned genes. AL-B alpha and beta contained 156 and 146 amino acid, respectively, including 23-residue signal peptides. AL-B beta showed the conserved hydrophilic patches, putative sites for GpIb binding. Furthermore, there was another conserved motif (SRTY) exclusively in platelet-agglutinating AL-B, TSV-GPIb-BP and Mamushigin. We propose that these three CLPs may function as bivalent adhesive proteins linking two GpIb molecules on adjacent platelets.
Arpijuntarangkoon
Jaradpong
J
Department of Medicine, Chulalongkorn University, Rama IV Road, Patumwan, Bangkok 10330, Thailand.
Rojnuckarin
Ponlapat
P
Muanpasitporn
Chuanchom
C
Kaeothip
Sophon
S
Sangvanich
Polkit
P
Intragumtornchai
Tanin
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry* / Q000235:genetics* / Q000494:pharmacology; D018076:DNA, Complementary; D006801:Humans; D037102:Lectins; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000037:antagonists & inhibitors; D017841:Trimeresurus
D003435:Crotalid Venoms; D018076:DNA, Complementary; D037102:Lectins; D019038:Platelet Glycoprotein GPIb-IX Complex; C112004:alboaggregin B