Article Sidebar
Published:
May 28, 2014Keywords:
Article Details
Submission of a manuscript to this journal is a representation that the manuscript has not been published previously and is not under consideration for publication elsewhere.
All authors named in each manuscript would be required to sign a form (to be supplied by the Editor) so that they may retain their copyright in the article but to assign to us (the Publishers) and its licensees in perpetuity, in all forms, formats and media (whether known or created in the future) to (i) publish, reproduce, distribute, display and store the contribution, (ii) translate the contribution into other languages, create adaptations, reprints, include within collections and create summaries, extracts and/or abstracts of the contribution, (iii) create any other derivative works(s) based on the contribution, (iv) to exploit all subsidiary rights in the contribution, (v) the inclusion of electronic links from the contribution to third party material where-ever it may be located, and (vi) license any thrid party to do any or all of the above.
Main Article Content
Isolation and Characterization of Antithrombin Peptides from the Saliva of Malaysian Leeches
Abstract
Purpose: To isolate and characterize the antithrombin compounds of Malaysian leeches’ saliva collection (LSC) for use as anticoagulant proteins and peptides.
Method: Reversed phase - high performance liquid chromatography (RP-HPLC) was used to isolate all proteins from LSC. All isolated proteins were tested for anticoagulant activity by applying two tests, thrombin time test and the inhibition of the amidolytic activity of thrombin on the chromogenic substrate (S-2238). The molecular weights of isolated active compounds were identified using tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE).
Results: More than 40 proteins were isolated from LSC using RP-HPLC. Two compounds (protein 1 and protein 2) were found to be active as they increased thrombin time by 30.26 and 36.75%, respectively and inhibited the amidolytic activity of thrombin. as evaluated by measuring the conversion of the chromogenic substrate (S-2238). Decrease in the conversion of the substrate (S-2238) by 31.10 and 41.61%, respectively was observed. The molecular weights of the proteins (6.289 and 14.255 kDa, respectively) had no similarity with other leech species.
Conclusion: Two antithrombin proteins with anticoagulant properties were isolated from the saliva of Malaysian leeches using one-step isolation procedure. The proteins did not show any similarity to previous isolated proteins from leech.
Keywords: Leech, Saliva, Antithrombin, Anticoagulant, Peptides, Molecular weight
