Abstract
Echistatin (Ech) is a potent inhibitor of integrins and was isolated from snake Echis carinatus. To facilitate the study on the molecular determinants of integrin–ligand interactions, we expressed recombinant Ech and its mutants in the Pichia pastoris (P. pastoris) expression system and purified them to homogeneity with the yields of 2–7 mg/L. Ech produced in P. pastoris inhibited platelet aggregation with the IC50 value of 210.5 nM. The sequential assignment and structure analysis of recombinant Ech were obtained by 2D and 3D 15N-edited NMR spectra. These data suggests that Ech produced in P. pastoris retained its function and native fold. The results presented here provide the evidences that four disulfide-bonded peptide inhibitor of integrin, Ech, can be expressed in P. pastoris with correct fold and high yield. Platelet aggregation analysis of Ech mutants showed that the length of C-terminus and the K45 residue of Ech are important for interacting with integrin αIIbβ3. We also found that recombinant Ech can inhibit the migration of human A375 melanoma cell. These findings may serve as the basis for understanding the activities of Ech.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
