In Vitro Study of the Fibrinolytic Activity via Single Chain Urokinase-Type Plasminogen Activator and Molecular Docking of FGFC1
Abstract
:1. Introduction
2. Results
2.1. Fibrinolytic Characterization of FGFC1
2.1.1. The Role of Pro-uPA in the Fibrinolytic Activity of FGFC1
2.1.2. The Role of Glu-Plasminogen in the Fibrinolytic Activity of FGFC1
2.1.3. The Fibrinolytic Characterization of FGFC1 Mediated by Pro-uPA and Glu-Plasminogen
2.1.4. The Fibrinolytic Characterization of FGFC1 Mediated by Pro-uPA and Lys-Plasminogen
2.2. Binding Sites and Mode of Binding between FGFC1 and Plasminogen
2.2.1. The Effect of EACA, TXA, and Soybean Trypsin Inhibitor (SBTI) on the Fibrinolytic Activity of FGFC1
2.2.2. Docking
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Fibrinolytic Characterization of FGFC1
- (Reaction 1): plasminogen activated to plasmin by pro-uPA;
- (Reaction 2): pro-uPA activated to uPA by plasmin;
- (Reaction 3): substrate S-2444 hydrolyzed by uPA.
4.2.1. The Role of Pro-uPA in the Fibrinolytic Activity of FGFC1
4.2.2. The Role of Glu-Plasminogen in the Fibrinolytic Activity of FGFC1
4.2.3. The FGFC1 Fibrinolytic Characterization by Pro-uPA and Glu-/Lys-Plasminogen
4.3. The Binding Sites and Binding Mode between FGFC1 and Plasminogen
4.3.1. The Effect of EACA, TXA, and SBTI on the Fibrinolytic Activity of FGFC1
4.3.2. Docking
- Step 1. Ligand Pretreatment
- Step 2. Receptor Pretreatment
- Step 3. Preparation of Docking Parameters
- Step 4. Running Molecular Docking and Output Results
- Step 5. Analysis of Docking Results for EACA and FGFC1 to KR1–KR5
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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FGFC1 | FGFC1 + EACA | FGFC1 + TXA | FGFC1 + SBTI | ||||
---|---|---|---|---|---|---|---|
Concentration (mM) | Fibrinolytic Activity (%) | Concentration of EACA (mM) | Fibrinolytic Activity (%) | Concentration of TXA (mM) | Fibrinolytic Activity (%) | Concentration of SBTI (mM) | Fibrinolytic Activity (%) |
0 | 100 | 0 | 100 | 0 | 100 | 0 | 100 |
0.048 | 139.36 ± 4.6 | 3.6 | 82.35 ± 8.7 | 0.72 | 86.19 ± 4.5 | 5 | 65.83 ± 1.2 |
0.072 | 210.64 ± 4.3 | 7.2 | 71.32 ± 1.5 | 1.08 | 64.55 ± 8.9 | 10 | 53.24 ± 1.5 |
0.096 | 218.09 ± 6.8 | 9.6 | 54.41 ± 5.4 | 1.44 | 51.87 ± 1.9 | 20 | 29.86 ± 3.9 |
0.12 | 207.45 ± 5.8 | 12 | 40.44 ± 0.1 | 2.4 | 23.88 ± 2.1 | 40 | 21.58 ± 2.3 |
0.18 | 134.04 ± 4.5 | 21.6 | 15.44 ± 1.9 | 4.8 | 8.21 ± 0.5 | 70 | 11.15 ± 0.6 |
0.24 | 98.94 ± 4.1 | 72 | 6.62 ± 2.3 | 14.4 | 4.10 ± 0.6 | 100 | 6.83 ± 2.6 |
0.36 | 74.47 ± 3.4 | 216 | 5.15 ± 2.5 | 21.6 | 3.36 ± 0.9 | 120 | 3.96 ± 0.5 |
Domain | EACA | FGFC1 | ||
---|---|---|---|---|
Residues in Hydrophobic Interactions | Residues in Hydrophilic Interactions | Residues in Hydrophobic Interactions | Residues in Hydrophilic Interactions | |
KR1 | Asp55, Asp57, Tyr64, Tyr72, Trp62 | Arg71, Arg35 | Pro58, Pro31, Trp62, Asp55, Arg35, Arg33, Tyr72, Lys70, Glu69 | His32, Arg71, Tyr64, Tyr74, Pro68, Asp57 |
KR2 | Asp54, Asp56, Trp61, Trp71, Phe63 | Arg70 | Trp71, Asp54, Tyr35, Lys43, Pro53, Asn52, Glu7, Asn42, Phe40, Asp56, Trp61 | Lys39, Asn55, Gly34 |
KR3 | Arg36, His33, His64, Trp72, Trp62 | Arg71, Lys57 | Asp81, Leu2, Val17, Lys76, Tyr74, Cys75, Glu73, Trp72, Ala60, Arg59, Thr5 | Ser79, Gly4, Ile 77 |
KR4 | Trp125, Trp135, Asp119, Asp121, Phe127 | Lys100, Arg134 | Ser92, Ser91, Pro95, Thr129, Cys87, Lys86, Asn113, Glu103, Met112, Met93, Gln88, Thr111, Thr101, Pro102 | Thr94, Ser89, Leu110 |
KR5 | Tyr72, Tyr74, Tyr64, Asp55, Asp57, Trp62, Phe36 | - | Asp57, Asp55, Tyr64, Tyr72, Leu71, Ile35, Trp62, Phe36, Thr40, Ser34 | Asn41, Arg32 |
Ligand | Receptor | Binding Affinity (kcal/mol) |
---|---|---|
EACA | KR1 | −5.2 |
EACA | KR2 | −4.3 |
EACA | KR3 | −3.7 |
EACA | KR4 | −4.5 |
EACA | KR5 | −4.3 |
FGFC1 | KR1 | −7.4 |
FGFC1 | KR2 | −9.0 |
FGFC1 | KR3 | −6.3 |
FGFC1 | KR4 | −8.3 |
FGFC1 | KR5 | −6.7 |
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Gao, C.; Shen, Q.; Tang, P.; Cao, Y.; Lin, H.; Li, B.; Sun, P.; Bao, B.; Wu, W. In Vitro Study of the Fibrinolytic Activity via Single Chain Urokinase-Type Plasminogen Activator and Molecular Docking of FGFC1. Molecules 2021, 26, 1816. https://doi.org/10.3390/molecules26071816
Gao C, Shen Q, Tang P, Cao Y, Lin H, Li B, Sun P, Bao B, Wu W. In Vitro Study of the Fibrinolytic Activity via Single Chain Urokinase-Type Plasminogen Activator and Molecular Docking of FGFC1. Molecules. 2021; 26(7):1816. https://doi.org/10.3390/molecules26071816
Chicago/Turabian StyleGao, Chunli, Quan Shen, Pengjie Tang, Yuling Cao, Houwen Lin, Bailin Li, Peng Sun, Bin Bao, and Wenhui Wu. 2021. "In Vitro Study of the Fibrinolytic Activity via Single Chain Urokinase-Type Plasminogen Activator and Molecular Docking of FGFC1" Molecules 26, no. 7: 1816. https://doi.org/10.3390/molecules26071816