1. High molecular weight kininogen inhibits fibrinogen binding to cytoadhesins of neutrophils and platelets
S Niewiarowski, Y T Wachtfogel, E J Gustafson, H Lukasiewicz, K J Norton, A H Schmaier, R W Colman J Cell Biol . 1989 Jul;109(1):377-87. doi: 10.1083/jcb.109.1.377.
Fibrinogen inhibited 125I-high molecular weight kininogen (HMWK) binding and displaced bound 125I-HMWK from neutrophils. Studies were performed to determine whether fibrinogen could bind to human neutrophils and to describe the HMWK-fibrinogen interaction on cellular surfaces. At 4 degrees C, the binding of 125I-fibrinogen to neutrophils reached a plateau by 30 min and did not decrease. At 23 and 37 degrees C, the amount of 125I-fibrinogen bound peaked by 4 min and then decreased over time because of proteolysis of fibrinogen by human neutrophil elastase (HNE). Zn++ (50 microM) was required for binding of 125I-fibrinogen to neutrophils at 4 degrees C and the addition of Ca++ (2 mM) increased the binding twofold. Excess unlabeled fibrinogen or HMWK completely inhibited binding of 125I-fibrinogen. Fibronectin degradation products (FNDP) partially inhibited binding, but prekallikrein and factor XII did not. The binding of 125I-fibrinogen at 4 degrees C was reversible with a 50-fold molar excess of fibrinogen or HMWK. Binding of 125I-fibrinogen, at a concentration range of 5-200 micrograms/ml of added radioligand, was saturable with an apparent Kd of 0.17 microM and 140,000 sites/cell. The binding of 125I-fibrinogen to neutrophils was not inhibited by the peptide RGDS derived from the alpha chain of fibrinogen or by the mAb 10E5 to the platelet glycoprotein IIb/IIIa heterodimer. Fibrinogen binding was inhibited by a gamma-chain peptide CYGHHLGGAKQAGDV and by mAb OKM1 but was not inhibited by OKM10, an mAb to a different domain of the adhesion glycoprotein Mac-1 (complement receptor type 3 [CR3]). HMWK binding to neutrophils was not inhibited by OKM1. These observations were consistent with a further finding that fibrinogen is a noncompetitive inhibitor of 125I-HMWK binding to neutrophils. Fibrinogen binding to ADP-stimulated platelets was increased twofold by Zn++ (50 microM) and was inhibited by HMWK. These studies indicate that fibrinogen specifically binds to the C3R receptor on the neutrophil surface through the carboxy terminal of the gamma-chain and that HMWK interferes with the binding of fibrinogen to integrins on both neutrophils and activated platelets.
2. PRFF Peptide Mimic Interferes with Toxic Fibrin-Aβ42 Interaction by Emulating the Aβ Binding Interface on Fibrinogen
Rajanya Bhattacharyya, Sayan Bhattacharjee ACS Chem Neurosci . 2021 Nov 3;12(21):4144-4152. doi: 10.1021/acschemneuro.1c00519.
Cerebrovascular dysfunction is a common phenomenon in Alzheimer's patients, where fibrinogen is a major player. With the blood-brain barrier compromised, fibrinogen gains access to the brain, where its interaction with Aβ42results in plasmin-resistant abnormal blood clots that are deposited in the cerebral blood vessels, a condition commonly encountered in Alzheimer's disease (AD) patients called cerebral amyloid angiopathy (CAA). So far, there have been no effective therapeutics available to combat AD-associated CAA. This study reports a 13-amino acid peptide (Pα-NPGRPEPGSAGTW) as a potential inhibitor of the fibrin-Aβ42interaction along with the property to dissolve pre-existing plasmin-resistant abnormal clots. Strikingly, the identified sequence was found to be partially similar to a fragment of the fibrinogen α-chain reported to bind Aβ42, the plasmin-resistant fibrinogen fragment (PRFF). Mechanistically, Pα interacts with Aβ42in place of fibrinogen, thus inhibiting the toxic fibrin-Aβ42interaction. However, it does not interfere with normal fibrin polymerization.
3. Inhibition of fibrinogen binding to human platelets by the tetrapeptide glycyl-L-prolyl-L-arginyl-L-proline
G Marguerie, E F Plow Proc Natl Acad Sci U S A . 1982 Jun;79(12):3711-5. doi: 10.1073/pnas.79.12.3711.
The role of fibrinogen as a cofactor in platelet aggregation is mediated by its binding to platelet receptors that are induced by stimuli such as ADP. In the present study, we demonstrate that the tetrapeptide glycyl-L-prolyl-L-arginyl-L-proline inhibits the interaction of fibrinogen with its platelet receptor. The primary effect of the peptide was on the extent rather than on the rate of fibrinogen binding. Significant inhibition occurred at a 1:1 molar ratio of peptide to fibrinogen and reached maximal levels at 100:1 ratio. The inhibition was dependent upon fibrinogen concentration and occurred in the presence of calcium or magnesium. The peptide inhibited the binding of fibrinogen to platelets with exposed receptors, suggesting that it interfered directly with the ligand-receptor interaction. Fibrinogen binding supported by epinephrine and thrombin as well as ADP was inhibited by the peptide. Fibrinogen-dependent aggregation of washed platelets by ADP was abolished by a 30-fold molar excess of the peptide. The tetrapeptide is an analog of the amino-terminal sequence of the alpha-chain of fibrin and has been shown to inhibit fibrin polymerization [Laudano, A. P. & Doolittle, R. F. (1978) Proc. Natl. Acad. Sci. USA 75, 3085-3089]. A peptide corresponding to the natural sequence, glycyl-L-prolyl-L-arginyl-L-valyl-L-valine, was also capable of inhibiting fibrinogen binding to the platelet. These results suggest that common structural features within fibrinogen may serve a dual function by permitting the molecule to participate in both platelet aggregation and fibrin formation.
