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A novel thrombin inhibitory peptide discovered from leech using affinity chromatography combined with ultra-high performance liquid chromatography-high resolution mass spectroscopy
Huang, Qiuyang1; Gao, Qian1; Chai, Xiaoxin1; Ren, Wei2; Zhang, Guifeng3; Kong, Yingjun3; Zhang, Yan3; Gao, Jianping3; Lei, Xiongxin3; Ma, Li1
2020-08-15
Source PublicationJOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
ISSN1570-0232
Volume1151Pages:8
Abstract

Thrombin (THR) inhibitors play an important role in the treatment of thrombotic diseases. This study established a THR-based bio-specific extraction coupled with affinity chromatography and ultra-high performance liquid chromatography-high resolution mass spectroscopy (UPLC-HR-MS) analysis method to screen and identify THR ligands in Leech. After evaluating the reliability of the screening method using positive control drug (hirudin), it was successfully used to screen the potential active constituents in leech. And a comprehensive analysis of the peptides in leech elution was performed by UPLC-HR-MS, a total of 34 peptides were identified. At the same time, anti-THR activity was explored and inferred by searching databases and published literature. As a result, six peptides were discovered to be potential active compounds in leech. Further, the six peptides were synthesized and in vitro enzymatic activity assay was performed. Finally, SYELPDGQVITIGNER was screened as an anti-THR peptide with an IC50, value of 255.75 mu M and it was discovered for the first time from Whitmania pigra Whitman and Hirudo nipponica Whitman. The molecular docking study showed that THR inhibitory activity of the polypeptide was mainly attributed to the hydrogen bond interactions, van der Waals forces and electrostatic interactions interaction between polypeptide and THR. These results suggest that the polypeptide is a potential natural THR inhibitor that can be used as anticoagulant.

KeywordThrombin Bioactive Compound Screening Affinity Chromatography Uplc-hr-ms Leech Thrombin Inhibitory Activity
DOI10.1016/j.jchromb.2020.122153
Language英语
WOS KeywordHirudin-like Factors ; Molecular Docking ; Medicinal Leech ; Ultrafiltration ; Hydrolysate ; Management ; Blood ; Drug
Funding ProjectNational Natural Science Foundation of China[81673596]
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
Funding OrganizationNational Natural Science Foundation of China
WOS IDWOS:000540214100001
PublisherELSEVIER
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/41104
Collection中国科学院过程工程研究所
Corresponding AuthorMa, Li
Affiliation1.Capital Med Univ, Sch Tradit Chinese Med, Beijing 100069, Peoples R China
2.Southwest Med Univ, Affiliated Tradit Chinese Med Hosp, Luzhou 646000, Peoples R China
3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
Recommended Citation
GB/T 7714
Huang, Qiuyang,Gao, Qian,Chai, Xiaoxin,et al. A novel thrombin inhibitory peptide discovered from leech using affinity chromatography combined with ultra-high performance liquid chromatography-high resolution mass spectroscopy[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2020,1151:8.
APA Huang, Qiuyang.,Gao, Qian.,Chai, Xiaoxin.,Ren, Wei.,Zhang, Guifeng.,...&Ma, Li.(2020).A novel thrombin inhibitory peptide discovered from leech using affinity chromatography combined with ultra-high performance liquid chromatography-high resolution mass spectroscopy.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,1151,8.
MLA Huang, Qiuyang,et al."A novel thrombin inhibitory peptide discovered from leech using affinity chromatography combined with ultra-high performance liquid chromatography-high resolution mass spectroscopy".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 1151(2020):8.
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