Related Experiment Video
Updated: Jul 9, 2025
Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation
Published on: May 23, 2025
Kenaf Seed Cysteine Protease (KSCP) Inhibits the Intrinsic Pathway of the Blood Coagulation Cascade and Platelet
Sujatha M Hanumegowda1, Chandramma Srinivasa2, Ashwini Shivaiah2
1Department of Biochemistry Jnansahydri, Kuvempu University, Shankarghatta-577451, Shivamogga, Karnataka, India.
Insights
Kenaf Seed Cysteine Protease (KSCP) exhibits significant antithrombotic potential by dissolving blood clots and inhibiting platelet aggregation. This natural compound offers a promising alternative for managing thrombotic complications without adverse effects.
Area of Science:
- Biochemistry
- Pharmacology
- Natural Product Chemistry
Background:
- Thrombosis poses a significant threat, causing stroke and myocardial infarction.
- Current antithrombotic drugs have severe side effects.
- Natural antithrombotic agents are gaining attention for their safety profile.
Purpose of the Study:
- To purify and characterize a cysteine protease from kenaf seed extract.
- To investigate the antithrombotic activity of the purified enzyme, Kenaf Seed Cysteine Protease (KSCP).
Main Methods:
- Purification using gel permeation and ion exchange chromatography.
- Characterization via SDS-PAGE, RP-HPLC, MALDI-TOF, and CD spectroscopy.
- Antithrombotic activity assessed through various coagulation and platelet aggregation assays.
Main Results:
- KSCP is a 24.1667 kDa cysteine protease with optimal activity at pH 6.0 and 40°C.
- KSCP demonstrated potent anticoagulant properties by inhibiting the intrinsic coagulation pathway.
- The enzyme effectively dissolved blood clots and inhibited platelet aggregation.
Conclusions:
- Kenaf Seed Cysteine Protease (KSCP) possesses significant antithrombotic potential.
- KSCP may serve as a valuable therapeutic candidate for managing thrombotic disorders.
- Further research into KSCP could lead to novel, safer antithrombotic therapies.
Background:
Thrombosis is the key event that obstructs the flow of blood throughout the circulatory system, leading to stroke, myocardial infarction and severe cardiovascular complications. Currently, available antithrombotic drugs trigger several life-threatening side effects.
Introduction:
Antithrombotic agents from natural sources devoid of adverse effects are grabbing high attention. In our previous study, we reported the antioxidant, anticoagulant and antiplatelet properties of kenaf seed protein extract. Therefore, in the current study, purification and characterization of cysteine protease from kenaf seed protein extract responsible for potential antithrombotic activity was undertaken.
Methods:
Purification of KSCP (Kenaf Seed Cysteine Protease) was carried out using gel permeation and ion exchange column chromatography. The purity of the enzyme was evaluated by SDS PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis). RP-HPLC (Reverse Phase High-Performance Liquid Chromatography), MALDI-TOF (Matrix-Assisted Laser Desorption Ionization Time-Of-Flight) and CD (Circular Dichroism techniques) were employed for its characterization. Proteolytic, fibrinolytic and kinetic study was done using spectroscopy. Plasma recalcification time, Prothrombin Time (PT), Thrombin clotting time (TCT), Activated Partial Thromboplastin Time (APTT), bleeding time and platelet aggregation studies were carried out for antithrombotic activity of KSCP.
Result:
A single sharp band of KSCP was observed under both reduced and non-reduced conditions, having a molecular mass of 24.1667kDa. KSCP was found to contain 30.3% helix turns and 69.7% random coils without a beta-pleated sheet. KSCP digested casein and fibrin, and its activity was inhibited by iodoacetic acid (IAA). KSCP was optimally active at pH 6.0 at the temperature of 40°C. KSCP exhibited anticoagulant properties by interfering in the intrinsic pathway of the blood coagulation cascade. Furthermore, KSCP dissolved both whole blood and plasma clots and platelet aggregation.
Conclusion:
KSCP purified from kenaf seed extract showed antithrombotic potential. Hence, it could be a better candidate for the management of thrombotic complications.
More Related Videos
07:13Author Spotlight: Developing Parmodulins to Target Protease-Activated Receptors for Inflammation Control
Published on: May 24, 2024
07:15Mechanism of Kemeng Fang's Inhibition of Podocyte Apoptosis in Rats with Membranous Nephropathy through the PI3K/AKT Signaling Pathway
Published on: August 23, 2024
Related Concept Videos
01:22Formation of the Platelet Plug
As the injured blood vessel contracts, endothelial cells undergo contraction, revealing collagen fibers in the basement membrane and underlying connective tissue. Furthermore, the plasma membrane of endothelial cells becomes adhesive, preparing the site for platelet adhesion. Platelets...
01:20Extrinsic and Intrinsic Pathways of Hemostasis
The Extrinsic Pathway
The extrinsic pathway of coagulation is typically initiated by tissue damage that exposes blood to tissue factor (TF), a protein released by the damaged tissue cells outside the blood vessels—this interaction with TF triggers biochemical reactions involving specific clotting factors. The key player here is Factor VII, which...
01:16Clot Retraction and Fibrinolysis
01:09Coagulation
During the coagulation phase, clotting factors, or procoagulants, play a vital role in initiating and progressing the coagulation cascade. This cascade is a series of reactions...
02:19The Spindle Assembly Checkpoint
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
01:05Introduction to Hemostasis
The three phases of hemostasis involve many clotting factors present in plasma and several substances released by platelets and injured tissue cells. It is a fast, localized,...