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Purification and characterization of Protein C activator from Agkistrodon acutus Venom.
Prep Biochem Biotechnol. 2020 Jun 01 [Online ahead of print]PB

Abstract

Protein C (PC) plays an important role in the balance of coagulation and anticoagulation. Thus, the detection of PC activity is diagnostically significant for patients with cardiovascular diseases. Presently, the key methods to detect PC activity are the chromogenic assay and activated partial thromboplastin time (APTT) test. PROTAC used in the chromogenic assay is isolated from Agkistrodon contortrix venom as protein C activator (PCA). However, the use of the chromogenic assay is limited because of the high price of PROTAC. In this study, PCA was successfully purified from Agkistrodon acutus venom (AAV) by ion-exchange and gel chromatography. PCA from AAV has a relative molecular mass of 24 kD, calculated from the measurement of 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The components of PCA were identified by MALDI-TOF/TOF-MS and mascot searches revealed that the coverage rate between PCA and zinc metalloproteinase AaPA from AAV was 21%. The chromogenic assay and APTT test were used to measure the enzymatic activity of PCA, and the results showed that PCA from AAV could specifically activate PC. In summary, the chromogenic assay described herein is highly sensitive and easy to perform.

Authors+Show Affiliations

Department of Pathophysiology, Institute of Snake Venom, Wannan Medical College, Wuhu, China.School of Anesthesiology, Wannan Medical College, Wuhu, China.Department of Pathophysiology, Institute of Snake Venom, Wannan Medical College, Wuhu, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32478599

Citation

Sun, Yao, et al. "Purification and Characterization of Protein C Activator From Agkistrodon Acutus Venom." Preparative Biochemistry & Biotechnology, 2020, pp. 1-8.
Sun Y, Bao PJ, Zhang GB. Purification and characterization of Protein C activator from Agkistrodon acutus Venom. Prep Biochem Biotechnol. 2020.
Sun, Y., Bao, P. J., & Zhang, G. B. (2020). Purification and characterization of Protein C activator from Agkistrodon acutus Venom. Preparative Biochemistry & Biotechnology, 1-8. https://doi.org/10.1080/10826068.2020.1768541
Sun Y, Bao PJ, Zhang GB. Purification and Characterization of Protein C Activator From Agkistrodon Acutus Venom. Prep Biochem Biotechnol. 2020 Jun 1;1-8. PubMed PMID: 32478599.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and characterization of Protein C activator from Agkistrodon acutus Venom. AU - Sun,Yao, AU - Bao,Peng-Ju, AU - Zhang,Gen-Bao, Y1 - 2020/06/01/ PY - 2020/6/2/pubmed PY - 2020/6/2/medline PY - 2020/6/2/entrez KW - Agkistrodon acutus venom KW - Protein C KW - Protein C activator SP - 1 EP - 8 JF - Preparative biochemistry & biotechnology JO - Prep. Biochem. Biotechnol. N2 - Protein C (PC) plays an important role in the balance of coagulation and anticoagulation. Thus, the detection of PC activity is diagnostically significant for patients with cardiovascular diseases. Presently, the key methods to detect PC activity are the chromogenic assay and activated partial thromboplastin time (APTT) test. PROTAC used in the chromogenic assay is isolated from Agkistrodon contortrix venom as protein C activator (PCA). However, the use of the chromogenic assay is limited because of the high price of PROTAC. In this study, PCA was successfully purified from Agkistrodon acutus venom (AAV) by ion-exchange and gel chromatography. PCA from AAV has a relative molecular mass of 24 kD, calculated from the measurement of 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The components of PCA were identified by MALDI-TOF/TOF-MS and mascot searches revealed that the coverage rate between PCA and zinc metalloproteinase AaPA from AAV was 21%. The chromogenic assay and APTT test were used to measure the enzymatic activity of PCA, and the results showed that PCA from AAV could specifically activate PC. In summary, the chromogenic assay described herein is highly sensitive and easy to perform. SN - 1532-2297 UR - https://www.unboundmedicine.com/medline/citation/32478599/Purification_and_characterization_of_Protein_C_activator_from_Agkistrodon_acutus_Venom L2 - http://www.tandfonline.com/doi/full/10.1080/10826068.2020.1768541 DB - PRIME DP - Unbound Medicine ER -
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