Abstract: A new in vitro anti-tumor polypeptide, coded as J2-C3, was isolated from Arca inflata Reeve and purified by diethyl-aminoethanol (DEAE)-sepharose Fast Flow anion exchange and phenyl sepharose CL-4B hydrophobic chromatography. J2-C3 was identified to be a homogeneous compound by native polyacrylamide gel electrophoresis (Native-PAGE). The purity of J2-C3 was over 99% in reversed phase-high performance liquid chromatography (RP-HPLC). The molecular weight was determined as 20,538.0 Da by electrospray-ionization mass spectrometry (ESI-MS/MS). J2-C3 was rich in Glx (Gln + Glu), Lys, and Asx (Asp + Asn) according to amino acid analysis. Four partial amino acid sequences of this peptide were determined as L/ISMEDVEESR, KNGMHSI/LDVNHDGR, AMKI/LI/LNPKKGI/LVPR and AMGAHKPPKGNEL/IGHR via MALDI-TOF/TOF-MS and de novo sequencing. Secondary structural analysis by CD spectroscopy revealed that J2-C3 had the α-helix (45.2%), β-sheet (2.9%), β-turn (26.0%) and random coil (25.9%). The anti-tumor effect of J2-C3 against human tumor cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the IC50 values of J2-C3 were 65.57, 93.33 and 122.95 µg/mL against A549, HT-29 and HepG2 cell lines, respectively. Therefore, J2-C3 might be developed as a potential anti-tumor agent.
Keywords: Arca inflata Reeve; polypeptide; structure analysis; anti-tumor activity
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Xu, J.; Chen, Z.; Song, L.; Chen, L.; Zhu, J.; Lv, S.; Yu, R. A New in Vitro Anti-Tumor Polypeptide Isolated from Arca inflata. Mar. Drugs 2013, 11, 4773-4787.
Xu J, Chen Z, Song L, Chen L, Zhu J, Lv S, Yu R. A New in Vitro Anti-Tumor Polypeptide Isolated from Arca inflata. Marine Drugs. 2013; 11(12):4773-4787.
Xu, Jian; Chen, Zhiyan; Song, Liyan; Chen, Lili; Zhu, Jianhua; Lv, Shuangshuang; Yu, Rongmin. 2013. "A New in Vitro Anti-Tumor Polypeptide Isolated from Arca inflata." Mar. Drugs 11, no. 12: 4773-4787.