title: Characterization of Collagen Peptides in Elaphuri Davidiani Cornu Aqueous Extract with Proliferative Activity on Osteoblasts Using Nano-Liquid Chromatography in Tandem with Orbitrap Mass Spectrometry.
abstract: First documented inShennong Bencao Jing(about 200 B.C.-200 A.D.),Elaphuri Davidiani Cornu(EDC) has been recorded for its effects in strengthening bones and balancing other aspects of overall health for approximately 2000 years. In the present study, our aim was to investigate which are the components of the active EDC fraction by a peptidomic strategy. We explored the extent to which EDC increases the proliferation of osteoblasts by measuring the elevations in osteonectin and type I collagen mRNA levels and characterized it using nano-flow liquid chromatography in tandem with orbitrap mass spectrometry. In total, 272 peptide sequences from collagens were determined. "Hot regions" in parent proteins determined by peptide heat maps which indicated that amino acid sequences in the regions might undergo proteolysis easily and generate peptides. Among the identified peptides, 90.2% were hydrophilic, and the molecular weight of 97.1% of identified peptides was lower than 2000 Da. According to these results, EDC collagen-derived peptides were easily analyzed and identified. Moreover, this methodology is feasible to characterize the active peptides matrices originated from collagen hydrolysates or some other animal horn- derived TCMs.
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. zhaiyanjuan1990@126.com.
authors.forename: Yanjuan
authors.initials: Y
authors.lastname: Zhai
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. 04040416@163.com.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. nzyzy808@163.com.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. qiandw@njucm.edu.cn.
authors.forename: Dawei
authors.initials: D
authors.lastname: Qian
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. cpulr@126.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.affiliation: Jiangsu Provincial Academy of Chinese Medicine, Nanjing 210028, China. pengyunru@126.com.
authors.forename: Yunru
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Jiangsu Dafeng Milu National Nature Reserve, Dafeng 224136, China. dingyuhuamilu@126.com.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Ding
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. zhenouyang@ujs.edu.cn.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Ouyang
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China. dja@njucm.edu.cn.
authors.forename: Jin-Ao
authors.initials: JA
authors.lastname: Duan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000090602:Collagen Type I, alpha 1 Chain; D003670:Deer; D006733:Horns / Q000737:chemistry*; D008516:Medicine, Chinese Traditional; D010006:Osteoblasts / Q000187:drug effects*; D015676:Osteonectin / Q000235:genetics / Q000378:metabolism*; D011492:Protein Hydrolysates / Q000494:pharmacology*; D051381:Rats; D053719:Tandem Mass Spectrometry; D014020:Tissue Extracts / Q000494:pharmacology*
chemical_list: D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D015676:Osteonectin; D011492:Protein Hydrolysates; D014020:Tissue Extracts
keywords: Elaphuri Davidiani Cornu; collagen peptide; nano-liquid chromatography-mass spectrum detection; osteoblast