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

• Published in: Protein and peptide letters Vol. 22; no. 5; p. 470
• Main Authors: Bao, Dongmei, Jin, Xiaobao, Ma, Yan, Zhu, Jiayong
• Format: Journal Article
• Language: English
• Published: Netherlands 01.01.2015
• Subjects: Amino Acid Sequence; Apoptosis - drug effects; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Cell Cycle - drug effects; Cell Movement - drug effects; Cell Proliferation - drug effects; Endostatins - chemistry; Endostatins - genetics; Endostatins - pharmacology; Hep G2 Cells; Humans; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver Neoplasms - drug therapy; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Models, Molecular; Molecular Sequence Data; Mutation; Peptides - chemistry; Peptides - genetics; Peptides - pharmacology; Protein Conformation; Protozoan Proteins - chemistry; Protozoan Proteins - genetics; Protozoan Proteins - pharmacology; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - pharmacology
• ISSN: 1875-5305
• DOI: 10.2174/0929866522666150302125218

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.