Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (15): 2390-2396.doi: 10.3969/j.issn.2095-4344.2014.15.016

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Primary structure of nuclease-like proteins from Eisenia foetida

Yu Bao-feng 1, 2, Liu Zhi-zhen 1, 2, Zhang Yue-hong 1, 2, Xie Jun 1, 2, Cheng Niu-liang 1, 2, Wang Jian-hua3, Niu Bo 1, 3    

  1. 1 Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 2 Key Laboratory of Cellular Physiology by Shanxi Province and State Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 3 Department of Biotechnology, Capital Institute of Pediatrics, Beijing 100020, China
  • Online:2014-04-09 Published:2014-04-09
  • Contact: Niu Bo, M.D., Professor, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Department of Biotechnology, Capital Institute of Pediatrics, Beijing 100020, China
  • About author:Yu Bao-feng, M.D., Associate professor, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Key Laboratory of Cellular Physiology by Shanxi Province and State Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 30472251; Youth Research Funds by the Natural Science Foundation of Shanxi Province, No. 2010021035-2; Shanxi Province Foundation for the Returned Overseas Scholars, No. 2010-52

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Abstract:

BACKGROUND: A group of nuclease-like proteins were previously purified from Eisenia foetida tissues, exploring primary structures of these proteins will help to uncover basic structure characteristics of them and provide foundations for the study addressing the relationship of their structures and functions.
OBJECTIVE: To explore primary structures of nuclease-like proteins EWD1 and EWD2.
METHODS: Edman degradation method was used to sequence the N-terminal amino acids of EWD1 and EWD2, acid hydrolisis method was used to analyze amino acid compositions of EWD1 and EWD2, LC-MS/MS was used to analyze some peptide sequences within the proteins, and MALDI-TOF-MS was used to calculate the number of the disulfide bonds and the contents of polysaccharides. 
RESULTS AND CONCLUSION: Among the amino acid compositions in EWD1 and EWD2, the sum contents of aspartate and asparagines were the highest (all nearly 10%), the contents of hydrophobic amino acids were also high, and the contents of cysteine was low. The EWD1 and EWD2 had similar amino acid compositions with other nucleases. Edman degradation results showed that, the N-terminal sequences of the large subunit of EWD1 were in turn as follows: D, E, W, V, Y, P; the N-terminal sequences of EWD2 were as follows: L, L, G, P, Y, K, P, K, C. The results of LC-MS/MS indicated the two proteins were novel proteins; MALDI-TOF-MS results showed that 8 cysteine residues formed 4 disulfide bonds in EWD1, 6 cysteine residues formed 3 disulfide bonds in EWD2. EWD1 and EWD2 were all glycoproteins, the content of polysaccharides was 17.3% in EWD1 and 15.6% in EWD2.



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


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Key words: oligochaeta, endonucleases, exonucleases, protein, amino acid sequence

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