The antiangiogenic and antitumor activities of the N-terminal fragment of endostatin augmented by Ile/Arg substitution: The overall structure implicated the biological activity.
The antiangiogenic and antitumor activities of the 27-amino acid fragment corresponding to the N-terminal domain of endostatin were shown to be dependent on a Zn-binding loop in the N-terminus. To investigate whether the regions outside of the N-terminal loop play a role in the peptide function, the structure and function of a variant containing Ile26Arg mutation (ES-R) were compared with those of the native peptide (ES-Zn). Structural analysis using far-UV CD, intrinsic fluorescence and molecular dynamics simulation provided information regarding the overall changes upon the mutation. In addition, the docking simulations predicted a higher affinity of ES-R to integrins αvβ3and α5β1than ES-Zn and a profound reorganization of the binding residues throughout the sequence. In Human Umbilical Vein Endothelial Cells (HUVECs), ES-R inhibited the tube formation and activated caspase-3 more strongly than do ES-Zn. Based on in vivo studies, the growth of breast tumor and expression of CD31, Bcl-2 and nonfunctional p53 were inhibited more effectively by ES-R than by ES-Zn. We conclude that the C-terminal region is involved in the peptide function through some global structural effects.
Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran.
Reyhane
R
Chamani
Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran. Electronic address: sm_asghari@guilan.ac.ir.
S Mohsen
SM
Asghari
Cancer Biology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Ali Mohammad
AM
Alizadeh
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Kamran
K
Mansouri
Shefa Neuroscience Research Center, Khatam Alanbia Hospital, Tehran, Iran.
Taher
T
Doroudi
Shefa Neuroscience Research Center, Khatam Alanbia Hospital, Tehran, Iran.
Peir Hossein
PH
Kolivand
Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran.
Hossein
H
Ghafouri
Department of Biology, Faculty of Science, Islamic Azad University, Parand, Iran.
Somayeh
S
Ehtesham
Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran.
Hanieh
H
Rabouti
Department of Life Sciences Engineering, Faculty of New Sciences & Technologies, University of Tehran, Iran.
Faramarz
F
Mehrnejad
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D020533:Angiogenesis Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D043169:Endostatins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000098:blood supply / Q000188:drug therapy / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D009389:Neovascularization, Pathologic / Q000517:prevention & control; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011487:Protein Conformation
D020533:Angiogenesis Inhibitors; D000970:Antineoplastic Agents; D043169:Endostatins; D010446:Peptide Fragments; C505407:CASP3 protein, human; D053148:Caspase 3
Angiogenesis; Apoptosis; Arginine; Endostatin peptide; Structure; Tumor