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Planta Med 2014; 80(11): 896-901
DOI: 10.1055/s-0034-1382836
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Purification and Characterization of a Novel Type I Ribosome Inactivating Protein, Pachyerosin, from Pachyrhizus erosus Seeds, and Preparation of its Immunotoxin against Human Hepatoma Cells

Jin-Lin Guo
1   Key Laboratory of Standardization of Chinese Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu, China
2   LuzhouLaoJiao Co., Ltd., Luzhou, China
,
Yuan-Liu Cheng
1   Key Laboratory of Standardization of Chinese Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu, China
,
Yi Qiu
1   Key Laboratory of Standardization of Chinese Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu, China
,
Cai-Hong Shen
2   LuzhouLaoJiao Co., Ltd., Luzhou, China
,
Bin Yi
2   LuzhouLaoJiao Co., Ltd., Luzhou, China
,
Cheng Peng
1   Key Laboratory of Standardization of Chinese Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu, China
› Author Affiliations
Further Information

Publication History

received 04 February 2014
revised 24 May 2014

accepted 04 June 2014

Publication Date:
16 July 2014 (online)

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Abstract

Pachyrhizus erosus seeds have a high protein content and are used in China due to their cytotoxic effect. Here we report the biological and pharmacological activity of the protein extracts from P. erosus seeds. A novel ribosome-inactivating protein, pachyerosin, from P. erosus seeds was successively purified to homogeneity using ammonium sulfate precipitation, DEAE-sepharose FF, and Sephacryl S-200. Pachyerosin showed to be a type I ribosome-inactivating protein with a molecular mass of 29 kDa and an isoelectric point of 9.19. It strongly inhibited protein synthesis of rabbit reticulocyte lysate with an IC50 of 0.37 ng/mL and showed N-glycosidase activity on rat liver ribosomes with an EC50 of 85.9 pM. The N-terminal 27 amino acids of pachyerosin revealed a 60.71 % sequence identity with abrin A from the seeds of Abrus precatorius. With the aim of targeting the delivery of pachyerosin, immunotoxin was prepared by conjugating pachyerosin with anti-human AFP monoclonal antibodies SM0736. The immunotoxin pachyerosin-SM0736 efficiently inhibited the growth of the human hepatoma cell line HuH-7 with an IC50 of 0.050 ± 0.004 nM, 2360 times lower than that of pachyerosin and 430 times lower than that of the immunotoxin against human gastric cancer cell line SGC7901. These results imply that pachyerosin may be used as a new promising anticancer agent.

Key words

Pachyrhizus erosus - Fabaceae - purification - ribosome-inactivating proteins - immunotoxin - anticancer agent
 

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