The Experts below are selected from a list of 141 Experts worldwide ranked by ideXlab platform
Charles W. Francis - One of the best experts on this subject based on the ideXlab platform.
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cell proliferation on fiBrin modulation By <B>FiBrinopeptideB> cleavage
Blood, 1995Co-Authors: Lee Ann Sporn, L A Bunce, Charles W. FrancisAbstract:FiBrin forms the cohesive network of hemostatic plugs and thromBi, and it also provides the temporary matrix for initial support of healing and revascularization. Because cell proliferation is needed for revascularization after vessel injury, we have characterized structural requirements of fiBrin needed to support cell proliferation on fiBrin in vitro. Proliferation of cultured human endothelial cells and fiBroBlasts was measured By 3H-thymidine incorporation on fiBrin surfaces varying in structure. FiBrin prepared with thromBin and lacking Both <B>FiBrinopeptideB>s A and B (desAB fiBrin) supported proliferation of Both endothelial cells and fiBroBlasts. In contrast, fiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A, supported significantly less proliferation. Also, fiBrin prepared By thromBin treatment of fiBrinogen lacking residues Beta 1–42 supported only a low level of proliferation. Therefore, <B>FiBrinopeptideB> B cleavage and exposure of Beta 15–42 enhanced proliferation of cells on fiBrin. Specific proteolytic inhiBitors were used to eliminate the potential mitogenic effects of residual fiBrin-Bound thromBin. Additional controls showed that neither catalytically inactive thromBin nor addition of the thromBin receptor-activating peptide (SFLLRNPNDKYEPF [SFLL]) stimulated proliferation on desA fiBrin. The results indicate that cell proliferation on fiBrin is enhanced By <B>FiBrinopeptideB> B cleavage and exposure of the amino terminus of the fiBrin Beta chain. They also show that specific structural features of the temporary fiBrin matrix formed at sites of injury may modulate the proliferative response of vascular cells.
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endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15 42 of the Beta chain
Journal of Clinical Investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:Adhesion and spreading ofcultured human umBilical vein endothelial cells on fiBrin surfaces ofvarying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role ofthe amino terminus of the fiBrin 6 chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B,) chain. After clotting with thromBin, this fiBrin derivative lacking B#1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion ofWeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 1542 of the fiBrin 8 chain. (J. Clin. Invest.
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Endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15-42 of the Beta chain.
The Journal of clinical investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:ABstract Adhesion and spreading of cultured human umBilical vein endothelial cells on fiBrin surfaces of varying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role of the amino terminus of the fiBrin Beta chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B Beta chain. After clotting with thromBin, this fiBrin derivative lacking B Beta 1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion of WeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 15-42 of the fiBrin Beta chain.
L A Bunce - One of the best experts on this subject based on the ideXlab platform.
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cell proliferation on fiBrin modulation By <B>FiBrinopeptideB> cleavage
Blood, 1995Co-Authors: Lee Ann Sporn, L A Bunce, Charles W. FrancisAbstract:FiBrin forms the cohesive network of hemostatic plugs and thromBi, and it also provides the temporary matrix for initial support of healing and revascularization. Because cell proliferation is needed for revascularization after vessel injury, we have characterized structural requirements of fiBrin needed to support cell proliferation on fiBrin in vitro. Proliferation of cultured human endothelial cells and fiBroBlasts was measured By 3H-thymidine incorporation on fiBrin surfaces varying in structure. FiBrin prepared with thromBin and lacking Both <B>FiBrinopeptideB>s A and B (desAB fiBrin) supported proliferation of Both endothelial cells and fiBroBlasts. In contrast, fiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A, supported significantly less proliferation. Also, fiBrin prepared By thromBin treatment of fiBrinogen lacking residues Beta 1–42 supported only a low level of proliferation. Therefore, <B>FiBrinopeptideB> B cleavage and exposure of Beta 15–42 enhanced proliferation of cells on fiBrin. Specific proteolytic inhiBitors were used to eliminate the potential mitogenic effects of residual fiBrin-Bound thromBin. Additional controls showed that neither catalytically inactive thromBin nor addition of the thromBin receptor-activating peptide (SFLLRNPNDKYEPF [SFLL]) stimulated proliferation on desA fiBrin. The results indicate that cell proliferation on fiBrin is enhanced By <B>FiBrinopeptideB> B cleavage and exposure of the amino terminus of the fiBrin Beta chain. They also show that specific structural features of the temporary fiBrin matrix formed at sites of injury may modulate the proliferative response of vascular cells.
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endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15 42 of the Beta chain
Journal of Clinical Investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:Adhesion and spreading ofcultured human umBilical vein endothelial cells on fiBrin surfaces ofvarying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role ofthe amino terminus of the fiBrin 6 chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B,) chain. After clotting with thromBin, this fiBrin derivative lacking B#1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion ofWeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 1542 of the fiBrin 8 chain. (J. Clin. Invest.
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Endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15-42 of the Beta chain.
The Journal of clinical investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:ABstract Adhesion and spreading of cultured human umBilical vein endothelial cells on fiBrin surfaces of varying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role of the amino terminus of the fiBrin Beta chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B Beta chain. After clotting with thromBin, this fiBrin derivative lacking B Beta 1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion of WeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 15-42 of the fiBrin Beta chain.
Carolyn J. Cassady - One of the best experts on this subject based on the ideXlab platform.
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the effects of trivalent lanthanide cationization on the electron transfer dissociation of acidic <B>FiBrinopeptideB> B and its analogs
Journal of the American Society for Mass Spectrometry, 2016Co-Authors: Juliette J Commodore, Carolyn J. CassadyAbstract:Electrospray ionization (ESI) on mixtures of acidic <B>FiBrinopeptideB> B and two peptide analogs with trivalent lanthanide salts generates [M + Met + H]4+, [M + Met]3+, and [M + Met –H]2+, where M = peptide and Met = metal (except radioactive promethium). These ions undergo extensive and highly efficient electron transfer dissociation (ETD) to form metallated and non-metallated c- and z-ions. All metal adducted product ions contain at least two acidic sites, which suggest attachment of the lanthanide cation at the side chains of one or more acidic residues. The three peptides undergo similar fragmentation. ETD on [M + Met + H]4+ leads to cleavage at every residue; the presence of Both a metal ion and an extra proton is very effective in promoting sequence-informative fragmentation. BackBone dissociation of [M + Met]3+ is also extensive, although cleavage does not always occur Between adjacent glutamic acid residues. For [M + Met – H ]2+, a more limited range of product ions form. All lanthanide metal peptide complexes display similar fragmentation except for europium (Eu). ETD on [M + Eu – H]2+ and [M + Eu]3+ yields a limited amount of peptide BackBone cleavage; however, [M + Eu + H]4+ dissociates extensively with cleavage at every residue. With the exception of the results for Eu(III), metallated peptide ion formation By ESI, ETD fragmentation efficiencies, and product ion formation are unaffected By the identity of the lanthanide cation. Adduction with trivalent lanthanide metal ions is a promising tool for sequence analysis of acidic peptides By ETD.
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dissociation of multiply charged negative ions for hirudin 54 65 <B>FiBrinopeptideB> B and insulin a oxidized
Journal of the American Society for Mass Spectrometry, 2001Co-Authors: Nigel P Ewing, Carolyn J. CassadyAbstract:Collision-induced dissociation (CID) was performed on multiply deprotonated ions from three commercial peptides: hirudin (54–65), <B>FiBrinopeptideB> B, and oxidized insulin chain A. Ions were produced By electrospray ionization in a Fourier transform ion cyclotron resonance mass spectrometer. Each of these peptides contains multiple acidic residues, which makes them very difficult to ionize in the positive mode. However, the peptides deprotonate readily making negative ion studies a viaBle alternative. The CID spectra indicated that the likely deprotonation sites are acidic residues (aspartic, glutamic, and cysteic acids) and the C-terminus. The spectra are rife with c, y, and internal ions, although some a, B, x, and z ions form. Many of the fragment ions were formed from cleavage adjacent to acidic residues, Both N- and C-terminal to the acidic site. In addition, neutral loss (e.g., NH3, CH3, H2O, and CO2) was prevalent from Both the parent ions and from fragment ions. These neutral eliminations were often indicative of specific amino acid residues. The fragmentation patterns from several charge states of the parent ions, when comBined, provide significant primary sequence information. These results suggest that negative mode CID of multiply deprotonated ions provides useful structural information and can Be worthwhile for highly acidic peptides that do not form positive ions in aBundance.
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negative ion matrix assisted laser desorption ionization time of flight post source decay caliBration By using <B>FiBrinopeptideB> B
Journal of the American Society for Mass Spectrometry, 1998Co-Authors: Jaran Jainhuknan, Carolyn J. CassadyAbstract:<B>FiBrinopeptideB> B (Mr 1552.58) was employed as a caliBration compound for matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) post-source decay (PSD) fragment ion analysis in the negative mode. Experiments were performed By using Both continuous and delayed extraction, with the maximum reflectron voltages Being 30 and 21 kV, respectively. For comparison, a common positive ion PSD caliBrant, ACTH(18–39) (Mr 2466.7), was also employed with positive ion caliBration constants Being applied to negative ion spectra. Using <B>FiBrinopeptideB> B as the caliBrant, the negative ion PSD results for angiotensin II (Mr 1046.2), renin suBstrate tetradecapeptide (horse) (Mr 1759.0), and the custom-synthesized peptide (K2G4)2 (Mr 987.1) showed a factor of 1.5–2 improvement in aBsolute mass accuracy. Typical aBsolute mass-to-charge ratio accuracies were within ±1 Thomson and were achieved even when the peptide Being analyzed was more massive than <B>FiBrinopeptideB> B. In addition, Both caliBrants showed increased accuracy when experiments were conducted in the delayed extraction mode. Other advantages of using <B>FiBrinopeptideB> B are its moderate cost and the aBility to perform caliBration and sample analysis for negative ion PSD under the same instrumental conditions.
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Negative Ion Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Post-Source Decay CaliBration By Using <B>FiBrinopeptideB> B
Journal of the American Society for Mass Spectrometry, 1998Co-Authors: Jaran Jai-nhuknan, Carolyn J. CassadyAbstract:<B>FiBrinopeptideB> B (Mr 1552.58) was employed as a caliBration compound for matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) post-source decay (PSD) fragment ion analysis in the negative mode. Experiments were performed By using Both continuous and delayed extraction, with the maximum reflectron voltages Being 30 and 21 kV, respectively. For comparison, a common positive ion PSD caliBrant, ACTH(18–39) (Mr 2466.7), was also employed with positive ion caliBration constants Being applied to negative ion spectra. Using <B>FiBrinopeptideB> B as the caliBrant, the negative ion PSD results for angiotensin II (Mr 1046.2), renin suBstrate tetradecapeptide (horse) (Mr 1759.0), and the custom-synthesized peptide (K2G4)2 (Mr 987.1) showed a factor of 1.5–2 improvement in aBsolute mass accuracy. Typical aBsolute mass-to-charge ratio accuracies were within ±1 Thomson and were achieved even when the peptide Being analyzed was more massive than <B>FiBrinopeptideB> B. In addition, Both caliBrants showed increased accuracy when experiments were conducted in the delayed extraction mode. Other advantages of using <B>FiBrinopeptideB> B are its moderate cost and the aBility to perform caliBration and sample analysis for negative ion PSD under the same instrumental conditions.
Craig A Pedersen - One of the best experts on this subject based on the ideXlab platform.
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urine and plasma levels of <B>FiBrinopeptideB> B in patients with deep vein thromBosis and pulmonary emBolism
Thrombosis Research, 2003Co-Authors: Timothy A Morris, James J Marsh, Craig M Burrows, Peter G Chiles, Ronald Konopka, Craig A PedersenAbstract:ABstract Background: Many patients with pulmonary thromBoemBolism remain undiagnosed, possiBly Because of the difficulty clinicians have in determining which patients merit work-up with accurate (But expensive) imaging techniques. OBjectives: We present the first prospective clinical study of pulmonary emBolism (PE) and deep venous thromBosis (DVT) detection using the FPBtot assay, which measures <B>FiBrinopeptideB> B and its first derivative, des-arginine <B>FiBrinopeptideB> B. Methods: Twenty three patients with signs or symptoms of PE or DVT were enrolled in the study prior to the performance of definitive testing. Using a novel immunoassay, FPBtot levels were measured in urine and plasma samples from patients as well as from healthy controls. Urine and plasma FPBtot levels were compared to the diagnostic results, as Blindly adjudicated By one of the investigators. Patients were excluded if they withdrew (n=1), had inconclusive diagnostic testing (n=7), or did not give samples (n=2 for urine, n=3 for plasma). Results: The mean FPBtot concentration in the urine of the ‘DVT/PE positive’ group was 78.4±35.2 ng/ml and 2.7±1.9 ng/ml in the ‘DVT/PE negative’ group (p=0.03). The urine FPBtot concentrations in the ‘DVT/PE negative’ group were not significantly different from those in the healthy control group (2.2±0.4 ng/ml, p=0.40). The area under the ROC curve for urine FPBtot concentrations was 97.3±3.8%, suggesting a high degree of diagnostic accuracy. Plasma FPBtot concentrations were not significantly different Between groups. Conclusions: Urine FPBtot levels may help detect patients with PE and DVT.
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Urine and plasma levels of <B>FiBrinopeptideB> B in patients with deep vein thromBosis and pulmonary emBolism.
Thrombosis research, 2003Co-Authors: Timothy A Morris, James J Marsh, Craig M Burrows, Peter G Chiles, Ronald G Konopka, Craig A PedersenAbstract:Many patients with pulmonary thromBoemBolism remain undiagnosed, possiBly Because of the difficulty clinicians have in determining which patients merit work-up with accurate (But expensive) imaging techniques. We present the first prospective clinical study of pulmonary emBolism (PE) and deep venous thromBosis (DVT) detection using the FPBtot assay, which measures <B>FiBrinopeptideB> B and its first derivative, des-arginine <B>FiBrinopeptideB> B. Twenty three patients with signs or symptoms of PE or DVT were enrolled in the study prior to the performance of definitive testing. Using a novel immunoassay, FPBtot levels were measured in urine and plasma samples from patients as well as from healthy controls. Urine and plasma FPBtot levels were compared to the diagnostic results, as Blindly adjudicated By one of the investigators. Patients were excluded if they withdrew (n =1), had inconclusive diagnostic testing (n = 7), or did not give samples (n = 2 for urine, n = 3 for plasma). The mean FPBtot concentration in the urine of the 'DVT/PE positive' group was 78.4 +/- 35.2 ng/ml and 2.7 +/- 1.9 ng/ml in the 'DVT/PE negative' group (p = 0.03). The urine FPB(tot) concentrations in the 'DVT/PE negative' group were not significantly different from those in the healthy control group (2.2 +/- 0.4 ng/ml, p = 0.40). The area under the ROC curve for urine FPB(tot) concentrations was 97.3 +/- 3.8%, suggesting a high degree of diagnostic accuracy. Plasma FPB(tot) concentrations were not significantly different Between groups. Urine FPBtot levels may help detect patients with PE and DVT.
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Urine and plasma levels of <B>FiBrinopeptideB> B in patients with deep vein thromBosis and pulmonary emBolism
Thrombosis Research, 2003Co-Authors: Timothy A Morris, James J Marsh, Craig M Burrows, Peter G Chiles, Ronald G Konopka, Craig A PedersenAbstract:ABstract Background: Many patients with pulmonary thromBoemBolism remain undiagnosed, possiBly Because of the difficulty clinicians have in determining which patients merit work-up with accurate (But expensive) imaging techniques. OBjectives: We present the first prospective clinical study of pulmonary emBolism (PE) and deep venous thromBosis (DVT) detection using the FPBtot assay, which measures <B>FiBrinopeptideB> B and its first derivative, des-arginine <B>FiBrinopeptideB> B. Methods: Twenty three patients with signs or symptoms of PE or DVT were enrolled in the study prior to the performance of definitive testing. Using a novel immunoassay, FPBtot levels were measured in urine and plasma samples from patients as well as from healthy controls. Urine and plasma FPBtot levels were compared to the diagnostic results, as Blindly adjudicated By one of the investigators. Patients were excluded if they withdrew (n=1), had inconclusive diagnostic testing (n=7), or did not give samples (n=2 for urine, n=3 for plasma). Results: The mean FPBtot concentration in the urine of the ‘DVT/PE positive’ group was 78.4±35.2 ng/ml and 2.7±1.9 ng/ml in the ‘DVT/PE negative’ group (p=0.03). The urine FPBtot concentrations in the ‘DVT/PE negative’ group were not significantly different from those in the healthy control group (2.2±0.4 ng/ml, p=0.40). The area under the ROC curve for urine FPBtot concentrations was 97.3±3.8%, suggesting a high degree of diagnostic accuracy. Plasma FPBtot concentrations were not significantly different Between groups. Conclusions: Urine FPBtot levels may help detect patients with PE and DVT.
Lee Ann Sporn - One of the best experts on this subject based on the ideXlab platform.
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cell proliferation on fiBrin modulation By <B>FiBrinopeptideB> cleavage
Blood, 1995Co-Authors: Lee Ann Sporn, L A Bunce, Charles W. FrancisAbstract:FiBrin forms the cohesive network of hemostatic plugs and thromBi, and it also provides the temporary matrix for initial support of healing and revascularization. Because cell proliferation is needed for revascularization after vessel injury, we have characterized structural requirements of fiBrin needed to support cell proliferation on fiBrin in vitro. Proliferation of cultured human endothelial cells and fiBroBlasts was measured By 3H-thymidine incorporation on fiBrin surfaces varying in structure. FiBrin prepared with thromBin and lacking Both <B>FiBrinopeptideB>s A and B (desAB fiBrin) supported proliferation of Both endothelial cells and fiBroBlasts. In contrast, fiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A, supported significantly less proliferation. Also, fiBrin prepared By thromBin treatment of fiBrinogen lacking residues Beta 1–42 supported only a low level of proliferation. Therefore, <B>FiBrinopeptideB> B cleavage and exposure of Beta 15–42 enhanced proliferation of cells on fiBrin. Specific proteolytic inhiBitors were used to eliminate the potential mitogenic effects of residual fiBrin-Bound thromBin. Additional controls showed that neither catalytically inactive thromBin nor addition of the thromBin receptor-activating peptide (SFLLRNPNDKYEPF [SFLL]) stimulated proliferation on desA fiBrin. The results indicate that cell proliferation on fiBrin is enhanced By <B>FiBrinopeptideB> B cleavage and exposure of the amino terminus of the fiBrin Beta chain. They also show that specific structural features of the temporary fiBrin matrix formed at sites of injury may modulate the proliferative response of vascular cells.
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endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15 42 of the Beta chain
Journal of Clinical Investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:Adhesion and spreading ofcultured human umBilical vein endothelial cells on fiBrin surfaces ofvarying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role ofthe amino terminus of the fiBrin 6 chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B,) chain. After clotting with thromBin, this fiBrin derivative lacking B#1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion ofWeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 1542 of the fiBrin 8 chain. (J. Clin. Invest.
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Endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and amino acid residues 15-42 of the Beta chain.
The Journal of clinical investigation, 1992Co-Authors: L A Bunce, Lee Ann Sporn, Charles W. FrancisAbstract:ABstract Adhesion and spreading of cultured human umBilical vein endothelial cells on fiBrin surfaces of varying structure were characterized to understand Better the interactions occurring Between endothelium and fiBrin at sites of vascular injury. FiBrin prepared with reptilase, which cleaves only <B>FiBrinopeptideB> A from fiBrinogen, and fiBrin prepared with thromBin, which cleaves Both <B>FiBrinopeptideB> A and <B>FiBrinopeptideB> B, equally supported endothelial cell adhesion. In contrast, only fiBrin made with thromBin mediated endothelial cell spreading, as assessed By fluorescence microscopy of cells stained with rhodamine phalloidin to identify actin stress fiBers or By scanning electron microscopy. FiBrin prepared with reptilase failed to support cell spreading. To further investigate the role of the amino terminus of the fiBrin Beta chain after <B>FiBrinopeptideB> B cleavage in promoting cell spreading, protease III from Crotalus atrox venom was used to specifically cleave the amino-terminal 42 residues of the fiBrinogen B Beta chain. After clotting with thromBin, this fiBrin derivative lacking B Beta 1-42 failed to support significant cell spreading. Spreading on fiBrin was unaffected By depletion of WeiBel-Palade Bodies from endothelial cells, indicating that the spreading was independent of stimulated von WilleBrand factor release. We conclude that endothelial cell spreading on fiBrin requires <B>FiBrinopeptideB> B cleavage and involves residues 15-42 of the fiBrin Beta chain.