Structural Insight into Integrin Recognition and Anticancer Activity of Echistatin.

ABSTRACT: Echistatin (Ech) is a short disintegrin with a long ⁴²NPHKGPAT C-terminal tail. We determined the 3-D structure of Ech by X-ray crystallography. Superimposition of the structures of chains A and B showed conformational differences in their RGD loops and C-termini. The chain A structure is consistent with our NMR analysis that the GPAT residues of the C-terminus cannot be observed due to high flexibility. The hydrogen bond patterns of the RGD loop and between the RGD loop and C-terminus in Ech were the same as those of the corresponding residues in medium disintegrins. The mutant with C-terminal HKGPAT truncation caused 6.4-, 7.0-, 11.7-, and 18.6-fold decreases in inhibiting integrins ?v?3, ?IIb?3, ?v?5, and ?5?1. Mutagenesis of the C-terminus showed that the H44A mutant caused 2.5- and 4.4-fold increases in inhibiting ?IIb?3 and ?5?1, and the K45A mutant caused a 2.6-fold decrease in inhibiting ?IIb?3. We found that Ech inhibited VEGF-induced HUVEC proliferation with an IC₅₀ value of 103.2 nM and inhibited the migration of A375, U373MG, and Panc-1 tumor cells with IC₅₀ values of 1.5, 5.7, and 154.5 nM. These findings suggest that Ech is a potential anticancer agent, and its C-terminal region can be optimized to improve its anticancer activity.

SUBMITTER: Chen YC 

PROVIDER: S-EPMC7695343 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature