Cloning of cDNAs and molecular characterisation of C-type lectin-like proteins from snake venoms

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Abstract

C-type lectin-like proteins (CTLPs) isolated from snake venoms are the largest and most complex non-mammalian vertebrate C-type lectin-like domain family. In the present study, we simultaneously amplified four cDNAs encoding different types of CTLP subunits from the venoms of two different species of snakes by RT-PCR with a single sense primer and a nested universal primer – two CTLP subunit-encoding cDNAs were cloned from Deinagkistrodon acutus venom and two from Agkistrodon halys Pallas venom. All four cloned CTLP subunits exhibited typical motifs in their corresponding domain regions but with relatively-low sequence similarities to each other. Compared with previously-published CTLPs, the four cloned CTLPs subunits showed slight variations in the calcium-binding sites and the disulphide bonding patterns. To our knowledge, these data constitute the first example of co-expression of CTLP platelet glycoprotein Ib-binding subunits and coagulation factors in Agkistrodon halys Pallas venom.

Highlights

▸ cDNA libraries prepared from lyophilised snake venoms. ▸ Four cDNAs encoding CTLP subunits were cloned from these using a single sense primer. ▸ Platelet glycoprotein Ib-binding subunit in the venom of Agkistrodon halys Pallas. ▸ CTLP subunits within a single venom targeting coagulation factors and platelet glycoproteins.

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