The Contribution of N-Terminal Region Residues of Cystatin A (Stefin A) to the Affinity and Kinetics of Inhibition of Papain, Cathepsin B, and Cathepsin L†

Biochemistry ◽  
1999 ◽  
Vol 38 (22) ◽  
pp. 7339-7345 ◽  
Author(s):  
Sergio Estrada ◽  
Alla Pavlova ◽  
Ingemar Björk
Keyword(s):  
Cathepsin B ◽  
Cathepsin L ◽  
Terminal Region ◽  
Stefin A ◽  
Cystatin A ◽  
Kinetics Of ◽  
Kinetics Of Inhibition

scholarly journals Cloning and Expression of the Major SecretedCathepsin B-Like Protein from Juvenile Fasciola hepatica andAnalysis of Immunogenicity following Liver FlukeInfection

2003 ◽  
Vol 71 (12) ◽  
pp. 6921-6932 ◽  
Author(s):  
Ruby H. P. Law ◽  
Peter M. Smooker ◽  
James A. Irving ◽  
David Piedrafita ◽  
Rebecca Ponting ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cathepsin B ◽  
Fasciola Hepatica ◽  
Cathepsin L ◽  
Cloning And Expression ◽  
Significant Similarity ◽  
Peptide Substrates ◽  
Liver Flukes ◽  
Kinetics Of

ABSTRACT The functions of the cathepsin B-like proteases in liver flukes are unknown and analysis has been hindered by a lack of protein for study, since the protein is produced in small amounts by juvenile flukes. To circumvent this, we isolated and characterized a cDNA encoding the major secreted cathepsin B from Fasciola hepatica. The predicted preproprotein is 339 amino acids in length, with the mature protease predicted to be 254 amino acids long, and shows significant similarity to parasite and mammalian cathepsin B. Only one of the two conserved histidine residues required for cathepsin B exopeptidase activity is predicted to be present. Recombinant preproprotein was produced in yeast, and it was shown that the recombinant proprotein can undergo a degree of self-processing in vitro to the mature form, which is active against gelatin and synthetic peptide substrates. The recombinant protein is antigenic in vaccinated rats, and antibodies to the protein are detected early after infection of rats and sheep with F. hepatica. The kinetics of the response to cathepsin B and cathepsin L after infection of sheep and rats confirm the temporal expression of these proteins during the life cycle of the parasite.

The Kinetics of Inhibition of the Action of Thrombin by the Fibrinopeptides Formed during the Clotting of Fibrinogen

1966 ◽  
Vol 16 (01/02) ◽  
pp. 277-295 ◽  
Author(s):  
A Silver ◽  
M Murray
Keyword(s):  
Amino Acids ◽  
Competitive Inhibition ◽  
Amorphous Material ◽  
Peptide Bond ◽  
Initial Reaction ◽  
Reaction Products ◽  
Coagulation Mechanism ◽  
Kinetics Of ◽  
Kinetics Of Inhibition ◽  
Burk Plot

SummaryVarious investigators have separated the coagulation products formed when fibrinogen is clotted with thrombin and identified fibrinopeptides A and B. Two other peaks are observed in the chromatogram of the products of coagulation, but these have mostly been dismissed by other workers. They have been identified by us as amino acids, smaller peptides and amorphous material (37). We have re-chromatographed these peaks and identified several amino acids. In a closed system of fibrinogen and thrombin, the only reaction products should be fibrin and peptide A and peptide B. This reasoning has come about because thrombin has been reported to be specific for the glycyl-arginyl peptide bond. It is suggested that thrombin also breaks other peptide linkages and the Peptide A and Peptide B are attacked by thrombin to yield proteolytic products. Thrombin is therefore probably not specific for the glycyl-arginyl bond but will react on other linkages as well.If the aforementioned is correct then the fibrinopeptides A and B would cause an inhibition with the coagulation mechanism itself. We have shown that an inhibition does occur. We suggest that there is an autoinhibition to the clotting mechanism that might be a control mechanism in the human body.The experiment was designed for coagulation to occur under controlled conditions of temperature and time. Purified reactants were used. We assembled an apparatus to record visually the speed of the initial reaction, the rate of the reaction, and the density of the final clot formed after a specific time.The figures we derived made available to us data whereby we could calculate and plot the information to show the mechanism and suggest that such an inhibition does exist and also further suggest that it might be competitive.In order to prove true competitive inhibition it is necessary to fulfill the criteria of the Lineweaver-Burk plot. This has been done. We have also satisfied other criteria of Dixon (29) and Bergman (31) that suggest true competitive inhibition.

scholarly journals Increased levels of the megakaryocyte and platelet expressed cysteine proteases stefin A and cystatin A prevent thrombosis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anna Mezzapesa ◽  
Delphine Bastelica ◽  
Lydie Crescence ◽  
Marjorie Poggi ◽  
Michel Grino ◽  
...  
Keyword(s):  
Cysteine Proteases ◽  
Stefin A ◽  
Cystatin A

scholarly journals The Conformation and the Aggregation Kinetics of α-Synuclein Depend on the Proline Residues in Its C-Terminal Region

Biochemistry ◽  
2010 ◽  
Vol 49 (43) ◽  
pp. 9345-9352 ◽  
Author(s):  
Jessika Meuvis ◽  
Melanie Gerard ◽  
Linda Desender ◽  
Veerle Baekelandt ◽  
Yves Engelborghs
Keyword(s):  
Terminal Region ◽  
Aggregation Kinetics ◽  
Kinetics Of
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