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从长牡蛎中分离得到的一种新型体外抗肿瘤多肽。

A new in vitro anti-tumor polypeptide isolated from Arca inflata.

机构信息

Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China.

出版信息

Mar Drugs. 2013 Dec 2;11(12):4773-87. doi: 10.3390/md11124773.

DOI:10.3390/md11124773
PMID:24317469
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3877886/
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.

摘要

从贻贝中分离得到一种新的体外抗肿瘤多肽 J2-C3，并通过二乙氨基乙醇（DEAE）-琼脂糖 Fast Flow 阴离子交换和苯基琼脂糖 CL-4B 疏水柱层析进行纯化。J2-C3 通过天然聚丙烯酰胺凝胶电泳（Native-PAGE）鉴定为均一化合物。J2-C3 在反相高效液相色谱（RP-HPLC）中的纯度超过 99%。电喷雾电离质谱（ESI-MS/MS）测定其分子量为 20538.0 Da。J2-C3 根据氨基酸分析富含 Glx（Gln+Glu）、Lys 和 Asx（Asp+Asn）。通过 MALDI-TOF/TOF-MS 和从头测序确定了该肽的四个部分氨基酸序列，分别为 L/ISMEDVEESR、KNGMHSI/LDVNHDGR、AMKI/LI/LNPKKGI/LVPR 和 AMGAHKPPKGNEL/IGHR。圆二色光谱法的二级结构分析表明 J2-C3 具有α-螺旋（45.2%）、β-折叠（2.9%）、β-转角（26.0%）和无规卷曲（25.9%）。J2-C3 对人肿瘤细胞的抗肿瘤作用通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定，J2-C3 对 A549、HT-29 和 HepG2 细胞系的 IC₅₀ 值分别为 65.57、93.33 和 122.95 µg/mL。因此，J2-C3 可能被开发为一种有潜力的抗肿瘤药物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/74e8ed0311fd/marinedrugs-11-04773-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/6cc134910c6b/marinedrugs-11-04773-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/b663ae874858/marinedrugs-11-04773-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/74e8ed0311fd/marinedrugs-11-04773-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/6cc134910c6b/marinedrugs-11-04773-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/b663ae874858/marinedrugs-11-04773-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/520a/3877886/74e8ed0311fd/marinedrugs-11-04773-g008.jpg

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