Comparison of the structure and biological activities of wild-type and mutant liver-targeting peptide modified recombinant human Endostatin (rES-CSP) in human hepatocellular carcinoma HepG2 cells

A missense mutation of aspartic to asparagine acid in 113 position of liver-targeting peptide csp i-plus modified rendostatin (res-csp) happened unexpectedly results in the changes of protein secondary structure and a reduced bioactivity. with the aim to clarify the structure-function relationships featuring the fuse protein res-csp,the three-dimensional structural models of wild-type and mutant d113nres-csp were constructed by template-based modeling approach. to evaluate the effect of the single mutation on res-csp stability,the molecular dynamic simulation was used to reveal the structural and dynamic characteristics. analysis on the bioactivity were conducted using a number of validated in vitro assays including proliferation,migration,cell cycle and apoptosis in hepg2 cells. results showed that the mutant res-csp reduce the stability and loss of function,and the wild-type res-csp could both bind to the normal liver cells chang's and the hepatoma cells hepg2 but significantly higher than non-targeted rendostatin. rescsp could inhibit the proliferation of hepatoma cells in a dose-dependent manner,and increase the proportion of g1 phase,reduce the proportion of s phase,promote the apoptosis on hepatoma cells. these results make a further complement of the mechanisms by which the fuse protein res-csp would provide a feasible and convenient approach to produce liver-targeting drugs for treatment of the liver disease. © 2015 bentham science publishers.