scholarly journals Catabolism of human tissue plasminogen activator in mice

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 539-544 ◽  
Author(s):  
HE Fuchs ◽  
H Jr Berger ◽  
SV Pizzo
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Active Site ◽  
Human Tissue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Organ Distribution ◽  
Tissue Plasminogen ◽  
Iodine 125 ◽  
Distribution Studies

The catabolism of human tissue plasminogen activator (t-PA) was studied in mice. The clearance of t-PA labeled with iodine 125 was rapid (t1/2). The clearance of phenylmethylsulfonyl-125I-t-PA, which is active site-inhibited, was identical to the active enzyme. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the vast majority of 125I-t-PA injected into the circulation was present as free enzyme and not in a complex with inhibitors. The clearance of 125I-t-PA was unaltered by large molar excesses of several ligands of known clearance specificities, including macroalbumin, asialoorosomucoid, and diisopropylphosphorylthrombin and was also not altered in the presence of a 1,000-fold molar excess of unlabeled t-PA. Organ distribution studies demonstrated that the early rapid clearance of 125I-t-PA occurred in hepatocytes, followed by a later renal phase of clearance. The clearance of 125I-urokinase (UK) also was studied and was very similar in all aspects to the clearance of 125I-t-PA. These results suggest that both t-PA and UK are cleared from the circulation by unique nonsaturable processes localized in the liver that are independent of the proteinase active site.

scholarly journals Catabolism of human tissue plasminogen activator in mice

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 539-544 ◽  
Author(s):  
HE Fuchs ◽  
H Jr Berger ◽  
SV Pizzo
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Active Site ◽  
Human Tissue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Organ Distribution ◽  
Tissue Plasminogen ◽  
Iodine 125 ◽  
Distribution Studies

Abstract The catabolism of human tissue plasminogen activator (t-PA) was studied in mice. The clearance of t-PA labeled with iodine 125 was rapid (t1/2). The clearance of phenylmethylsulfonyl-125I-t-PA, which is active site-inhibited, was identical to the active enzyme. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the vast majority of 125I-t-PA injected into the circulation was present as free enzyme and not in a complex with inhibitors. The clearance of 125I-t-PA was unaltered by large molar excesses of several ligands of known clearance specificities, including macroalbumin, asialoorosomucoid, and diisopropylphosphorylthrombin and was also not altered in the presence of a 1,000-fold molar excess of unlabeled t-PA. Organ distribution studies demonstrated that the early rapid clearance of 125I-t-PA occurred in hepatocytes, followed by a later renal phase of clearance. The clearance of 125I-urokinase (UK) also was studied and was very similar in all aspects to the clearance of 125I-t-PA. These results suggest that both t-PA and UK are cleared from the circulation by unique nonsaturable processes localized in the liver that are independent of the proteinase active site.

scholarly journals Expression of Active Recombinant Human Tissue-Type Plasminogen Activator by Using In Vivo Polyhydroxybutyrate Granule Display

2010 ◽  
Vol 76 (21) ◽  
pp. 7226-7230 ◽  
Author(s):  
Yanping Geng ◽  
Shengjun Wang ◽  
Qingsheng Qi
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Human Tissue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Surface Display ◽  
Sodium Dodecyl ◽  
Tissue Type ◽  
Affinity Tag ◽  
Tissue Plasminogen

ABSTRACT Recombinant human tissue plasminogen activator (rPA) is a truncated version of tissue plasminogen activator (tPA), which contains nine disulfide bonds and is prone to forming inactive inclusion bodies when expressed in bacteria. To obtain functional rPA expression, we displayed the rPA on the surface of polyhydroxybutyrate (PHB) granules using phasin as the affinity tag. rPA was fused to the N terminus of the phasin protein with a thrombin cleavage site as the linker. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis showed that rPA fusion was successfully displayed on the surface of PHB granules. An activity assay indicated that the rPA fusion is active. The in vivo surface display strategy for functional rPA expression in Escherichia coli is distinct for its efficient folding and easier purification and may be expanded to the expression of other eukaryotic proteins with complex conformation.

Cloning and Expression of a Human Tissue Plasminogen Activator Variant:K2S in Escherichia coli

2007 ◽  
Vol 10 (6) ◽  
pp. 946-949 ◽  
Author(s):  
Hamid Mir Mohammad S ◽  
Kianoush Dormiani ◽  
Yahya Khazaie ◽  
Mohammad Rabbani ◽  
Fatemeh Moazen
Keyword(s):  
Escherichia Coli ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Human Tissue ◽  
Cloning And Expression ◽  
Tissue Plasminogen

Inhibitors Interfering with the Quantitative Estimation of Tissue Plasminogen Activator

1975 ◽  
Author(s):  
G. Wijngaards ◽  
A. T. Potjer
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Human Tissue ◽  
Inhibitory Action ◽  
Human Lung ◽  
Quantitative Estimation ◽  
Plasminogen Activators ◽  
Tissue Extracts ◽  
Tissue Plasminogen ◽  
Inhibitory Components

The commonly used quantitative assay for plasminogen activator in tissues by Astrup and Albrechtsen (1957) was re-evaluated with special reference to the presence of inhibitors in tissue extracts at different stages of the procedure.Human lung, liver, and placenta tissues were chosen for their different activator and inhibitor content. Samples of the extracts were tested for inhibitory capacity against purified human tissue plasminogen activator, urokinase, and plasmin on different kinds of fibrin plates.The procedure for preparing the samples according to Astrup and Albrechtsen did not completely eliminate the inhibitory action against the plasminogen activators added. The presence of inhibitors was partly obscured by 2M KCNS in the sample. In order to quantitate the plasminogen activator content in tissues, the method should be revised with respect to the elimination of inhibiting material.Some interesting aspects of the inhibitory components extracted from liver and placenta were revealed by comparing urokinase inhibition to tissue activator inhibition as to the degree of denaturation during the procedure. It was suggested that, although they have much in common, urokinase and tissue activator inhibitors are separate entities.

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